Pseudomonas sp. Pse-1 and application thereof

Abstract

The present application relates to a kind of pseudomonas Pse-1 and its application.A kind of pseudomonas, its preservation number is CGMCC No.22583.The above-mentioned pseudomonas is applied in oil production in oil production, in microbial oil production, in the application of producing oil bacteria liquid.The beneficial effects of the present application are that (1) the pseudomonas Pse-1 provided in the present application, its growth temperature is 37~45 ℃, can be degenerated from the fluid without viscosity under reservoir condition to a kind of multifunctional oil production bacteria liquid with surface activity and viscoelasticity, also has the function of expanding swept area and washing oil, physical simulation experiment improves recovery more than 20%;(2) the fermentation broth of the pseudomonas Pse-1 is applied to microbial single well throughput, average single well oil increment is greater than 500 tons, effective period is more than 250d;(3) the fermentation broth of the pseudomonas Pse-1 is applied to microbial flooding, well group average daily oil increment is more than 5 tons.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of energy biotechnology and environmental biotechnology, and particularly relates to a Pseudomonas sp. Pse-1 and application thereof. BACKGROUND

[0002] Two-thirds of the crude oil in most of the domestic major oilfields remains in the reservoir and cannot be produced after the application of conventional oil production technology, but the oilfields have entered the high water cut development stage. How to improve the recovery efficiency of the oilfields in the high water cut development stage is the key to the stable production of old oilfields.

[0003] Microbial oil production technology uses injected or activated microorganisms with oil displacement function in the reservoir to improve the oil recovery rate through the interaction between the metabolic activities of the microorganisms and the metabolic products thereof and the crude oil and the reservoir. A large number of indoor researches and field tests have been carried out at home and abroad, although the technology shows good application prospects, but it has not been realized large-scale popularization and application in the field.

[0004] The development of the reservoir is complex, and the oil displacement efficiency and the expanded sweep need to be solved at the same time to effectively improve the oil recovery rate. The existing microbial oil production technology mainly uses activated endogenous microbial flora or injected exogenous strains. The complexity of the endogenous microbial flora makes it difficult to accurately activate and control the oil target flora, and the exogenous bacteria have the limitation of single function, which cannot effectively solve the various contradictions in the reservoir.

[0005] The strain is the material basis and key of the microbial oil production technology, and obtaining a new type of multifunctional and efficient exogenous strain, as well as solving the expanded sweep system and the oil displacement efficiency are the keys to effectively improve the efficiency of the microbial oil production technology. SUMMARY

[0006] The first object of the present application is to provide a Pseudomonas sp. The second object of the present application is to provide the application of the Pseudomonas sp.

[0007] Technical scheme: A Pseudomonas sp. (Pseudomonas sp.) Pse-1, the preservation number of which is CGMCC No. 22583.

[0008] A microbial agent, the effective component of which is the Pseudomonas sp.

[0009] Application of the substance I and / or the substance II and / or the substance III and / or the substance IV and / or the substance V in oil production;

[0010] The substance I is the Pseudomonas sp. mentioned above;

[0011] The substance II is the microbial agent mentioned above;

[0012] The substance III is the bacterial suspension of the Pseudomonas sp. mentioned above;

[0013] Substance IV is a culture broth of the above-mentioned Pseudomonas;

[0014] Substance V is a fermentation product of the above-mentioned Pseudomonas.

[0015] Use of substance I and / or substance II and / or substance III and / or substance IV and / or substance V as a microbial agent in oil recovery;

[0016] Substance I is the above-mentioned Pseudomonas;

[0017] Substance II is the above-mentioned bacterial agent;

[0018] Substance III is a bacterial suspension of the above-mentioned Pseudomonas;

[0019] Substance IV is a culture broth of the above-mentioned Pseudomonas;

[0020] Substance V is a fermentation product of the above-mentioned Pseudomonas.

[0021] Use of substance I and / or substance II and / or substance III and / or substance IV and / or substance V in microbial oil recovery;

[0022] Substance I is the above-mentioned Pseudomonas;

[0023] Substance II is the above-mentioned bacterial agent;

[0024] Substance III is a bacterial suspension of the above-mentioned Pseudomonas;

[0025] Substance IV is a culture broth of the above-mentioned Pseudomonas;

[0026] Substance V is a fermentation product of the above-mentioned Pseudomonas.

[0027] Use of substance I and / or substance II and / or substance III and / or substance IV and / or substance V in the production of an oil recovery bacterial broth, which comprises a temperature-sensitive viscoelastic biological polysaccharide and a biological surfactant;

[0028] Substance I is the above-mentioned Pseudomonas;

[0029] Substance II is the above-mentioned bacterial agent;

[0030] Substance III is a bacterial suspension of the above-mentioned Pseudomonas;

[0031] Substance IV is a culture broth of the above-mentioned Pseudomonas;

[0032] Substance V is a fermentation product of the above-mentioned Pseudomonas.

[0033] Further, in the application, the Pseudomonas is produced in the nutrient medium with glycerol, xylose as carbon source, sodium nitrate and yeast powder as nitrogen source, dipotassium hydrogen phosphate as phosphorus source, adding inorganic salt sodium chloride, and containing trace elements magnesium and calcium.

[0034] Further, in the application, the nutrient medium comprises: xylose 8-12 g / L, glycerol 15-35 g / L, sodium nitrate 1-4 g / L, dipotassium hydrogen phosphate 1-3 g / L, sodium chloride 1-3 g / L, calcium chloride 0.05-0.1 g / L, magnesium sulfate 0.3-0.7 g / L, yeast powder 0.5-2 g / L, pH 7.5-8.0, and the balance is water.

[0035] Preferably, the nutrient medium comprises: xylose 10 g / L, glycerol 30 g / L, sodium nitrate 2 g / L, dipotassium hydrogen phosphate 2 g / L, sodium chloride 2 g / L, calcium chloride 0.1 g / L, magnesium sulfate 0.5 g / L, yeast powder 1 g / L, pH 7.8, and the balance is water.

[0036] Further, in the application, the fermentation conditions of the Pseudomonas are as follows: inoculum 1%-3%, inoculation age 12-24 h, initial pH 7.5-8.0, temperature 37-45℃, stirring speed 160-220 rpm, aeration amount 1.0-2.0 L / min, and fermentation time 48-72 h.

[0037] Preferably, the fermentation conditions of the Pseudomonas are as follows: inoculum 2%, inoculation age 24 h, initial pH 7.5, temperature 37℃, stirring speed 180 rpm, aeration amount 1.5 L / min, and fermentation time 60 h.

[0038] An oil production bacterial liquid, which comprises a temperature-sensitive viscoelastic biological polysaccharide and a biological surfactant, and the active substance in the oil production bacterial liquid is substance I and / or substance II and / or substance III and / or substance IV and / or substance V.

[0039] The substance I is the Pseudomonas as described above.

[0040] The substance II is the bacterial agent as described above.

[0041] The substance III is the bacterial suspension of the Pseudomonas as described above.

[0042] The substance IV is the culture solution of the Pseudomonas as described above.

[0043] The substance V is the fermentation product of the Pseudomonas as described above.

[0044] The function of the oil production bacterial liquid is (a) or (b) or (c) as follows:

[0045] (a) oil production;

[0046] (b) microbial oil recovery;

[0047] (c) producing the microbial oil recovery bacteria solution.

[0048] A method for oil recovery, comprising the step of adding substance I and / or substance II and / or substance III and / or substance IV and / or substance V during the oil recovery process.

[0049] Substance I is the Pseudomonas described above.

[0050] Substance II is the bacterial agent described above.

[0051] Substance III is the bacterial suspension of the Pseudomonas described above.

[0052] Substance IV is the culture solution of the Pseudomonas described above.

[0053] Substance V is the fermentation product of the Pseudomonas described above.

[0054] A method for producing a microbial oil recovery bacteria solution, which comprises thermosensitive viscoelastic biological polysaccharide and biological surfactant, comprising the steps of:

[0055] The microbial oil recovery bacteria solution is produced by fermentation culture of the Pseudomonas described above in a nutrient medium.

[0056] The inoculation amount of the Pseudomonas is 1% to 3%, the inoculation age is 12 to 24 hours, the initial pH is 7.5 to 8.0, the temperature is 37 to 45°C, the stirring speed is 160 to 220 rpm, the aeration amount is 1.0 to 2.0 L / min, and the fermentation time is 48 to 72 hours.

[0057] The nutrient medium is: xylose 8 to 12 g / L, glycerol 15 to 35 g / L, sodium nitrate 1 to 4 g / L, dipotassium hydrogen phosphate 1 to 3 g / L, sodium chloride 1 to 3 g / L, calcium chloride 0.05 to 0.1 g / L, magnesium sulfate 0.3 to 0.7 g / L, yeast powder 0.5 to 2 g / L, pH 7.5 to 8.0, and the balance is water.

[0058] Application of substance I and / or substance II and / or substance III and / or substance IV and / or substance V in microbial single-well huff and puff and / or microbial oil displacement.

[0059] Substance I is the Pseudomonas described above.

[0060] Substance II is the bacterial agent described above.

[0061] Substance III is the bacterial suspension of the Pseudomonas described above.

[0062] Substance IV is the culture solution of the Pseudomonas described above.

[0063] Substance V is the fermentation product of the Pseudomonas described above.

[0064] Furthermore, in the aforementioned applications, the microbial single-well huff and puff and / or microbial flooding are applicable to reservoirs with reservoir temperatures of 60–95°C and crude oil dehydration and degassing viscosity ≤10000 mPa·s at 50°C.

[0065] Furthermore, in the aforementioned application, the microbial single-well huff and puff involves injecting Pseudomonas fermentation broth into the formation through an oil well, utilizing the synergistic effect of biosurfactants and temperature-sensitive viscoelastic biopolysaccharides produced by Pseudomonas to improve single-well productivity.

[0066] Furthermore, in the aforementioned application, the specific steps of the single-well microbial huff and puff process are as follows:

[0067] Add 200-500 ml of Pseudomonas fermentation broth with a concentration of 30%-50%. 3 Injected into the formation via the annulus through the casing, with an injection rate of 10–20 m / s. 3 / h, the well is shut in for cultivation for 10-30 days before being put back into production, utilizing the synergistic effect of biosurfactants and temperature-sensitive viscoelastic polysaccharides to improve single-well productivity.

[0068] Furthermore, in the aforementioned application, the formula for calculating the injection volume of Pseudomonas fermentation broth is as follows:

[0069] V = 3.14R 2 Hфβ, where:

[0070] V—Volume of Pseudomonas fermentation broth injected, in m³ 3 ;

[0071] R—processing radius, in meters;

[0072] H—Effective thickness, in meters (m);

[0073] ф—porosity;

[0074] β—Dosage coefficient, with a value of 1.0 to 1.5.

[0075] Furthermore, in the aforementioned application, the microbial enhanced oil recovery involves injecting Pseudomonas fermentation broth into the water injection section via a plug injection method, utilizing the metabolic products of the Pseudomonas fermentation broth to improve the production capacity of the well group.

[0076] Furthermore, in the aforementioned application, the microbial enhanced oil recovery method involves injecting the Pseudomonas fermentation broth from the injection well using an alternating slug injection technique at an injection rate of 30–60 m / s. 3 / d, injection volume per round 100-200m 3 The specific steps include:

[0077] The Pseudomonas fermentation liquor with a concentration of 30%-50% is injected from a water injection well, normal water injection is carried out for 5-7 days, then water injection is stopped for 1-2 days, and then normal water injection is carried out for 10-14 days, so that a cycle of injection is completed.

[0078] The advantages and beneficial effects of the present application are that:

[0079] (1) The Pseudomonas Pse-1 provided by the present application has a growth temperature of 37-45 DEG C, can be denatured from a fluid without viscosity into a multifunctional oil recovery bacterial liquid with surface activity and viscoelasticity under oil reservoir conditions, has the functions of expanding swept area and oil washing, the viscosity of the multifunctional oil recovery bacterial liquid after denaturation of the temperature-sensitive biological polysaccharide is greater than 50 mPa s, the surface tension of the bacterial liquid is less than 30 mN / m, the oil washing efficiency is greater than 90%, and the physical simulation experiment improves the recovery rate by more than 20%.

[0080] (2) The fermentation liquor of the Pseudomonas Pse-1 is applied to microbial single-well huff and puff, and the average single-well oil increment is greater than 500 tons, and the effective period is more than 250 days;

[0081] (3) The fermentation liquor of the Pseudomonas Pse-1 is applied to microbial oil displacement, and the average daily oil increment of a well group is more than 5 tons. BRIEF DESCRIPTION OF DRAWINGS

[0082] Figure 1 It is a phylogenetic tree diagram of the Pseudomonas Pse-1 disclosed in the present application;

[0083] Figure 2 It is a colony morphology diagram of the Pseudomonas Pse-1 disclosed in the present application;

[0084] Figure 3 It is a morphology diagram of the Pseudomonas Pse-1 after staining of the bacterial body disclosed in the present application;

[0085] Figure 4 It is a morphology diagram of the network-like polysaccharide structure in the bacterial liquid after denaturation of the Pseudomonas Pse-1 at 60 DEG C disclosed in the present application

[0086] Figure 5 It is a viscosity change diagram of the Pseudomonas Pse-1 fermentation liquor at different times at 60 DEG C disclosed in the present application;

[0087] Biological material preservation for patent procedures:

[0088] Preservation date: May 24, 2021

[0089] Preservation unit: China General Microbiological Culture Collection Center

[0090] Address of deposit: No. 3, Yikuangli, Beichenxi Road, Beijing, China Institute of Microbiology, Chinese Academy of Sciences

[0091] Deposit number: CGMCC No. 22583

[0092] Classification name: Pseudomonas sp. DETAILED DESCRIPTION

[0093] The endpoints of the ranges and any values disclosed in this application are not limited to the precise values recited as the exact range or value should be understood as being approximated by the values in the range or value. For ranges of values, the endpoints of each range are inclusive of the values in the range, and the endpoints of each range are individually combinable with the endpoints of other ranges to form new ranges of values that are not expressly disclosed.

[0094] The nutrient medium (nutrient solution) used in the following specific examples is as follows: xylose 8-12 g / L, glycerol 15-35 g / L, sodium nitrate 1-4 g / L, potassium phosphate dibasic 1-3 g / L, sodium chloride 1-3 g / L, calcium chloride 0.05-0.1 g / L, magnesium sulfate 0.3-0.7 g / L, yeast powder 0.5-2 g / L, pH 7.5-8.0, and the balance is water.

[0095] The preferred scheme of the nutrient medium is as follows: xylose 10 g / L, glycerol 30 g / L, sodium nitrate 2 g / L, potassium phosphate dibasic 2 g / L, sodium chloride 2 g / L, calcium chloride 0.1 g / L, magnesium sulfate 0.5 g / L, yeast powder 1 g / L, pH 7.8, and the balance is water.

[0096] The specific embodiments of the present application are described in detail below.

[0097] Example 1

[0098] Obtaining and identification of Pseudomonas Pse-1 of the present application

[0099] I. Obtaining of Pseudomonas Pse-1

[0100] Pseudomonas Pse-1 of the present application is isolated from the produced fluid of an oil well in a certain block of Shengli Oilfield, and its classification name is Pseudomonas sp., abbreviated as Pse-1.

[0101] II. Identification of Pseudomonas Pse-1

[0102] 16s rDNA amplification and sequencing are performed on Pse-1 for strain identification, and the steps are as follows: denaturation, PCR amplification, PCR product purification, and sequencing of the target fragment. According to the sequencing results, sequence alignment is performed in NCBI, and a phylogenetic tree is constructed.Figure 1 ), identified the Pseudomonas Pse-1 of the present application as a new bacteria, belonging to the genus of Pseudomonas.

[0103] According to the experimental method of "Bergey's Mannual of Systematic Bacteriology", the size and morphology of the bacteria, the presence or absence of flagella and spores, the growth temperature, the aerobic condition, and the colony morphology.

[0104] The research results show that, Figure 2 and Figure 3 As shown in FIGS. 1 and 2, the strain has a short rod shape, a single flagellum, and can move, the colony is raised, the edge is uneven, the surface is smooth and moist, and the colony surface has sticky and filamentous polysaccharide.

[0105] The cell is long rod-shaped, and the size is (0.1-0.2) x (2-3) μm.

[0106] The physiological and biochemical characteristics of Pseudomonas Pse-1: aerobic, growth temperature 37-45℃, optimum growth temperature 37℃, growth pH range 5-10, optimum growth pH range 7.8-8, NaCl tolerance 0-10%.

[0107] Example 2

[0108] Performance evaluation parameters of Pseudomonas Pse-1:

[0109] Surface tension of fermentation broth under reservoir conditions, oil sand washing performance of fermentation broth, surface viscosity, and physical simulation experiment of enhanced oil recovery value.

[0110] Performance evaluation method of Pseudomonas Pse-1:

[0111] (1) Surface tension and apparent viscosity of fermentation broth under different dilution multiples under reservoir conditions

[0112] Pseudomonas Pse-1 was inoculated in a nutrient medium at an inoculation amount of 2%, and was incubated at 37℃ for 60h. The fermentation broth was diluted, heated at the reservoir temperature for 8h, and then the surface tension and apparent viscosity of the fermentation broth at different dilution degrees were detected. The properties of the viscous fermentation broth are shown in FIGS. 3 and 4, and the specific test data are shown in the following table: Figure 4 Figure 5

[0113] Dilution Surface tension (mN / m) Apparent viscosity (mPa-s) 10％ 24.7 150 20％ 25.2 121 30％ 25.4 107 40％ 26.7 86 50％ 28.3 65 60％ 35.7 24

[0114] (2) Performance evaluation of fermentation broth for washing oil sand

[0115] According to the standard "Q / SH10201518-2013", oil sand was prepared and a standard curve was drawn. 3.0g of oil sand was weighed into a 50mL colorimetric tube, and was marked as A.​​

[0116] The Pse-1 fermentation broth was heated at reservoir temperature for 8 h, and then diluted at different ratios.

[0117] 10 mL of the treated and diluted Pse-1 fermentation broth was added to a colorimetric tube, which was covered and placed in a 60°C constant-temperature water bath. The colorimetric tube was taken out every 15 min, shaken gently 10 times, and then placed in the water bath again. After 1 h, the colorimetric tube was taken out.

[0118] The upper liquid was carefully poured out, and the residual bacterial liquid in the colorimetric tube was repeatedly washed with distilled water until the washing liquid was transparent. The upper liquid was carefully poured out.

[0119] Another 50 mL colorimetric tube was taken, labeled as B, and placed on the colorimetric tube rack. The funnel was inserted into the colorimetric tube.

[0120] The colorimetric tube A was placed directly above the funnel, with the opening inclined downward. The bottom of A was washed with a washing bottle filled with distilled water. The oil sand in A was carefully transferred to B, and the upper washing liquid was poured out.

[0121] The colorimetric tube with oil sand was placed in a 105°C oven for drying for 4 h, and then taken out and placed in a desiccator until room temperature. The colorimetric tube with oil sand was added with an appropriate amount of petroleum ether with a boiling range of 60-90°C, shaken thoroughly, and diluted to the scale. The petroleum ether solution was taken and the absorbance was measured on a separate photometer. The residual oil content in the colorimetric tube was determined on a standard curve. The oil washing rate X = (1-W1 / KW0) x 100%, wherein W1 is the residual oil content of the oil sand in the colorimetric tube, K is the mass fraction of oil in the oil sand, and W0 is the mass of the oil sand.

[0122] The oil washing efficiency X = (1-W1 / KW0) x 100% = (1-0.0046 / 0.02 x 3.0067) x 100% = 92.4%. The oil washing efficiency of the fermentation broth under different dilutions was finally evaluated, and the results are shown in the following table:

[0123] Dilution Oil washing efficiency (%) 10％ 96.4 20％ 95.2 30％ 93.1 40％ 92.3 50％ 91.5 60％ 84.2

[0124] (4) Physical simulation experiment to evaluate the enhanced oil recovery value

[0125] ① Core preparation: core packing and sterilization, and determination of air permeability;

[0126] ② Vacuumization and saturation with simulated formation water, and determination of core PV (pore volume);

[0127] ③ Saturation with crude oil, core aging for 7 d, and calculation of irreducible water saturation;

[0128] ④ Primary water flooding, water flooding until the water produced contains more than 98% water, and calculation of primary water flooding recovery rate;

[0129] (5) Inject 0.2-0.4 PV of Pseudomonas Pse-1 fermentation broth, close the injection end and the production end, and denature at the reservoir temperature for 8-10 hours;

[0130] (6) Secondary water flooding, water flooding to the production liquid containing 100% water, calculate the Pseudomonas Pse-1 enhanced recovery value, reaching 23%.

[0131] The performance of Pseudomonas Pse-1: 30%-50% of the fermentation broth has a surface tension of less than 30 mN / m, and a viscosity of more than 50 mN / m after denaturation; the oil washing efficiency is more than 90%; the physical simulation experiment enhanced recovery is more than 20%.

[0132] Example 3

[0133] Application of Pseudomonas Pse-1 as an oil production bacterium liquid.

[0134] In the above application, the Pseudomonas is produced in a nutrient medium with glycerol, xylose as carbon source, sodium nitrate and yeast powder as nitrogen source, dipotassium hydrogen phosphate as phosphorus source, adding inorganic salt sodium chloride, and containing trace elements magnesium and calcium to produce the multifunctional oil production bacterium liquid.

[0135] The nutrient medium is xylose 8-12 g / L, glycerol 15-35 g / L, sodium nitrate 1-4 g / L, dipotassium hydrogen phosphate 1-3 g / L, sodium chloride 1-3 g / L, calcium chloride 0.05-0.1 g / L, magnesium sulfate 0.3-0.7 g / L, yeast powder 0.5-2 g / L, and pH is 7.5-8.0.

[0136] In the first example, the nutrient medium is xylose 10 g / L, glycerol 30 g / L, sodium nitrate 2 g / L, dipotassium hydrogen phosphate 2 g / L, sodium chloride 2 g / L, calcium chloride 0.1 g / L, magnesium sulfate 0.5 g / L, yeast powder 1 g / L, and pH is 7.8.

[0137] In the second example, the nutrient medium is xylose 12 g / L, glycerol 35 g / L, sodium nitrate 4 g / L, dipotassium hydrogen phosphate 3 g / L, sodium chloride 3 g / L, calcium chloride 0.1 g / L, magnesium sulfate 0.7 g / L, yeast powder 2 g / L, and pH is 7.5, and the rest is water.

[0138] In the third example, the nutrient medium is xylose 8 g / L, glycerol 15 g / L, sodium nitrate 1 g / L, dipotassium hydrogen phosphate 1 g / L, sodium chloride 1 g / L, calcium chloride 0.05 g / L, magnesium sulfate 0.3 g / L, yeast powder 0.5 g / L, and pH is 8.0, and the rest is water.

[0139] In this embodiment, the fermentation conditions of the Pseudomonas were as follows: inoculum size 2%, inoculum age 12h, initial pH 7.5, temperature 37℃, stirring speed 220rpm, aeration rate 1L / min, and fermentation time 72h.

[0140] In another embodiment, the fermentation conditions of the Pseudomonas were as follows: inoculum size 3%, inoculum age 24h, initial pH 7.5, temperature 40℃, stirring speed 200rpm, aeration rate 1.5L / min, and fermentation time 60h.

[0141] In the third embodiment, the fermentation conditions of the Pseudomonas were as follows: inoculum size 1%, inoculum age 12h, initial pH 8.0, temperature 45℃, stirring speed 160rpm, aeration rate 2.0L / min, and fermentation time 48h.

[0142] A multifunctional oil-producing bacterial solution is obtained by fermentation culture of the aforementioned Pseudomonas bacteria.

[0143] Example 4

[0144] The application of the Pseudomonas Pse-1 fermentation broth provided by this invention in a heavy oil well A1 in a certain block of Shengli Oilfield.

[0145] Oil well overview: Formation temperature 65℃, formation water salinity 18712 mg / L, effective oil layer thickness 6.7 m, porosity 35%, permeability 567 × 10⁻⁶. -3 μm 2 The viscosity of crude oil at 50℃ is 3764 mPa·s.

[0146] Application of the above-mentioned Pseudomonas Pse-1 fermentation broth in microbial single-well huff and puff.

[0147] Furthermore, the microbial single-well huff and puff involves injecting Pseudomonas Pse-1 fermentation broth into the formation through an oil well, utilizing the synergistic effect of biosurfactants and temperature-sensitive viscoelastic polysaccharides to improve single-well productivity.

[0148] Furthermore, the specific steps of the aforementioned microbial single-well throughput are as follows:

[0149] Use 200-500 ml of the prepared Pseudomonas Pse-1 fermentation broth with a concentration of 30%-50% 3 Injected into the formation via the annulus of the oil well casing at an injection rate of 10–20 m / s. 3 / h, the well is shut in for cultivation for 10-30 days before being put back into production, utilizing the synergistic effect of biosurfactants and temperature-sensitive viscoelastic polysaccharides to improve the productivity of a single well.

[0150] The specific implementation steps are as follows: Calculate the injection volume of fermentation fluid based on the effective thickness, porosity, and treatment radius of the oil layer in the oil well.

[0151] Injection total amount formula: V = 3.14R 2 Hβ, wherein

[0152] V - injection amount, m 3 ;

[0153] R - treatment radius, 7 m;

[0154] H - effective thickness (6.7 m);

[0155] φ - porosity (take 0.35);

[0156] β - amount coefficient (take 1.1),

[0157] According to the formula, the amount of Pseudomonas Pse-1 fermentation broth is 397 m 3 ; the prepared Pse-1 fermentation broth is injected into the formation through the oil well casing annulus by a pump truck; the oil well is closed for 30 days and then opened for production.

[0158] Test results: after the well A1 resumes production, the liquid production and oil production are both increased, the average daily oil increase of a single well reaches 2.8 t, the cumulative oil increase is 980 t, the minimum crude oil viscosity is reduced to 762 mPa·s, the reduction reaches 80%, the effective period reaches 350 d, and the field test effect is good.

[0159] Example 5

[0160] The application provides the application of Pseudomonas Pse-1 fermentation broth in a heavy oil well B1 in a certain block of Shengli Oilfield.

[0161] Oil well profile: formation temperature 60℃, formation water salinity 22140 mg / L, oil layer effective thickness 5.1 m, porosity 30%, permeability 467 x 10 -3 μm 2 , crude oil viscosity 1875 mPa·s.

[0162] The application of the above-mentioned Pseudomonas Pse-1 fermentation broth in microbial single-well stimulation.

[0163] Further, the microbial single-well stimulation is to inject Pseudomonas Pse-1 fermentation broth into the formation through the oil well, and to improve the single-well productivity by the synergistic effect of the biosurfactant and the temperature-sensitive viscoelastic biological polysaccharide.

[0164] The specific implementation steps are as follows: the injection amount of the fermentation broth is calculated according to the oil layer effective thickness, porosity and treatment radius of the oil well.

[0165] Injection total amount formula: V = 3.14R 2 Hβ, wherein

[0166] V - injection amount, m 3 ;

[0167] R - treatment radius, 6 m;

[0168] H - effective thickness (5.1 m);

[0169] f - porosity (take 0.30);

[0170] β - amount coefficient (take 1.2),

[0171] According to the formula, the amount of Pseudomonas Pse-1 fermentation broth is 208 m 3 ; the prepared Pse-1 fermentation broth is injected into the formation through the oil well casing annulus by a pump truck; the oil well is closed for 20 days and then opened for production.

[0172] Test results: after the well B1 resumes production, the liquid production and oil production are both increased, the average daily oil increase of single well reaches 3.7 t, the cumulative oil increase is 1166 t, the minimum crude oil viscosity is reduced to 283 mPa·s, the reduction reaches 84.9%, the effective period reaches 315 days, and the field test effect is good.

[0173] Example 6

[0174] The application provides the application of Pseudomonas Pse-1 fermentation broth in a certain block C1 of Shengli Oilfield, and the well group 1 is injected and 5 is extracted.

[0175] Oil well profile: formation temperature 65℃, formation water salinity 24134 mg / L, oil layer effective thickness 4.7 m, porosity 32%, permeability 653*10 -3 μm 2 , crude oil viscosity 2719 mPa·s.

[0176] The above-mentioned application of Pseudomonas Pse-1 fermentation broth in microbial oil displacement.

[0177] Further, the microbial oil displacement is to inject the Pseudomonas fermentation broth from the water injection well in a slug injection mode, and to improve the productivity of the well group by using the metabolic products of the Pseudomonas Pse-1 fermentation broth.

[0178] Further, the microbial displacement adopts a slug injection mode, the injection speed is 30 m 3 / d, the injection amount of each round is 100 m 3 , and the specific steps include:

[0179] The Pseudomonas Pse-1 fermentation broth with a concentration of 30% is injected from the water injection well for 3 days, normal water injection is performed for 5 days, then the injection is stopped for 1 day, and then normal water injection is performed for 13 days, to complete one cycle of injection.

[0180] Test results: A total of 5 injections were performed in this well group. After injection, the production of fluid and oil in all wells increased. The average daily oil production per well reached 6.3t, with a cumulative increase of 13,261t. The crude oil viscosity dropped to a minimum of 316mPa·s, a reduction of 88%, and the effective period reached 421 days. The field test results were good.

[0181] Example 7

[0182] The application of the Pseudomonas Pse-1 fermentation broth provided by this invention in Block D1 of Shengli Oilfield, where the well group has 1 injection and 4 production.

[0183] Well Overview: Formation temperature 72℃, formation water salinity 19473 mg / L, effective oil layer thickness 5.7 m, porosity 35%, permeability 429 × 10⁻⁶. -3 μm 2 The crude oil viscosity is 3271 mPa·s.

[0184] Application of the above-mentioned Pseudomonas Pse-1 fermentation broth in microbial oil recovery.

[0185] Furthermore, the microbial enhanced oil recovery involves injecting Pseudomonas fermentation broth into the water injection section via a plug injection method, utilizing the metabolites of the Pseudomonas fermentation broth Pse-1 to improve the production capacity of the well group.

[0186] Furthermore, the microbial flooding method employs alternating slug injection at a rate of 50m. 3 / d, injection volume per round 150m 3 The specific steps include:

[0187] The 40% Pseudomonas Pse-1 fermentation broth was injected from the injection well for 3 days, followed by 6 days of normal water injection, then stopped for 2 days, and then injected normally for 14 days to complete one cycle of injection.

[0188] Test results: The well group was injected in a total of 7 rounds. After injection, the production of fluid and oil in all wells increased. The average daily oil production per well reached 5.5t, with a cumulative increase of 8316t. The crude oil viscosity dropped to a minimum of 215mPa·s, a reduction of 93%, and the effective period reached 378 days. The field test results were good.

[0189] The preferred embodiments of the present invention have been described in detail above; however, the present invention is not limited thereto. Within the scope of the inventive concept, various simple modifications can be made to the technical solutions of the present invention, including combinations of various technical features in any other suitable manner. These simple modifications and combinations should also be considered as the content disclosed in the present invention and are all within the protection scope of the present invention.

Claims

1. Use of substance I and / or substance II and / or substance III and / or substance IV in the production of oil recovery bacteria liquid, wherein the oil recovery bacteria liquid comprises a temperature-sensitive viscoelastic biological polysaccharide and a biological surfactant; the substance I is a Pseudomonas sp. Pse-1, and the preservation number is CGMCC No. 22583; the substance II is a bacterial agent, and the effective component is a Pseudomonas sp. Pse-1, and the preservation number is CGMCC No. 22583; the substance III is a bacterial suspension of the substance I; the substance IV is a culture solution of the substance I, wherein: the Pseudomonas sp. is produced in a nutrient medium with glycerol and xylose as carbon sources, sodium nitrate and yeast powder as nitrogen sources, dipotassium hydrogen phosphate as a phosphorus source, and the addition of inorganic salt sodium chloride and trace elements magnesium and calcium. The fermentation conditions of the Pseudomonas sp. are as follows: inoculum 1% to 3%, inoculation age 12 to 24 hours, initial pH 7.5 to 8.0, temperature 37 to 45°C, stirring speed 160 to 220 rpm, aeration amount 1.0 to 2.0 L / min, and fermentation time 48 to 72 hours. The nutrient medium comprises: xylose 8 to 12 g / L, glycerol 15 to 35 g / L, sodium nitrate 1 to 4 g / L, dipotassium hydrogen phosphate 1 to 3 g / L, sodium chloride 1 to 3 g / L, calcium chloride 0.05 to 0.1 g / L, magnesium sulfate 0.3 to 0.7 g / L, yeast powder 0.5 to 2 g / L, and pH 7.5 to 8.0, and the balance is water. The nutrient medium comprises: xylose 10 g / L, glycerol 30 g / L, sodium nitrate 2 g / L, dipotassium hydrogen phosphate 2 g / L, sodium chloride 2 g / L, calcium chloride 0.1 g / L, magnesium sulfate 0.5 g / L, yeast powder 1 g / L, and pH 7.8, and the balance is water. The fermentation conditions of the Pseudomonas sp. are as follows: inoculum 2%, inoculation age 24 hours, initial pH 7.5, temperature 37°C, stirring speed 180 rpm, aeration amount 1.5 L / min, and fermentation time 60 hours. The method comprises the following steps: The oil recovery bacteria liquid is produced by fermentation culture of a Pseudomonas sp. Pse-1 with a preservation number of CGMCC No. 22583 in a nutrient medium; 2. Use according to claim 1, wherein The inoculum of the Pseudomonas sp. is 1% to 3%, the inoculation age is 12 to 24 hours, the initial pH is 7.5 to 8.0, the temperature is 37 to 45°C, the stirring speed is 160 to 220 rpm, the aeration amount is 1.0 to 2.0 L / min, and the fermentation time is 48 to 72 hours.

3. Use according to claim 2, wherein the compound is ###0002### The nutrient medium comprises: xylose 8 to 12 g / L, glycerol 15 to 35 g / L, sodium nitrate 1 to 4 g / L, dipotassium hydrogen phosphate 1 to 3 g / L, sodium chloride 1 to 3 g / L, calcium chloride 0.05 to 0.1 g / L, magnesium sulfate 0.3 to 0.7 g / L, yeast powder 0.5 to 2 g / L, and pH 7.5 to 8.0, and the balance is water.

4. The use according to claim 1, wherein 6. Use of substance I and / or substance II and / or substance III and / or substance IV in microbial single-well huff and puff and / or microbial oil displacement.

5. A method for preparing an oil recovery bacterial broth comprising a temperature-sensitive viscoelastic biopolymer and a biosurfactant, characterized in that, ​ ​ ​ ​ ​ The substance I is a Pseudomonas sp. Pse-1, and its preservation number is CGMCC No. 22583; The substance II is a microbial agent, and its effective component is a Pseudomonas sp. Pse-1, and its preservation number is CGMCC No. 22583; The substance III is a microbial suspension of the substance I; The substance IV is a culture solution of the substance I, and wherein: The microbial single-well huff and puff and / or microbial oil displacement are applicable to oil reservoirs with a temperature of 60-95 DEG C and a crude oil dehydration and degassing viscosity of less than or equal to 10000 mPa s at 50 DEG C; The microbial single-well huff and puff is achieved by injecting a Pseudomonas fermentation broth into a formation through an oil well, and the specific steps are as follows: Pseudomonas fermentation broth with concentration of 30% to 50% 200 to 500 m 3 The oil jacket annulus is injected into the stratum at an injection speed of 10 to 20 m 3 / h, and the oil well is shut down for 10 to 30 days before being opened for production, thereby improving the single well productivity by the synergistic effect of the biological surfactant and the temperature-sensitive viscoelastic biological polysaccharide.

7. Use according to claim 6, wherein The injection amount of the Pseudomonas fermentation broth is calculated according to the following formula: V = 3.14R 2 Hβ, where: V - injection amount of Pseudomonas fermenting liquid, unit: m 3 ; R is a treatment radius, and the unit is m; H is an effective thickness, and the unit is m; φ is porosity; β is a dosage coefficient, and the value is 1.0-1.

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8. The use according to claim 6, wherein The microbial oil displacement is achieved by injecting a Pseudomonas fermentation broth from a water injection well in a slug mode, and the metabolic products of the Pseudomonas fermentation broth are used to improve the productivity of the well group.

9. The use according to claim 6, wherein The microbial oil displacement is to inject the Pseudomonas fermentation broth from the water injection well by the way of slug alternating injection, the injection speed is 30-60 m 3 / d, the injection amount of each round is 100-200 m 3 , the specific steps include: The Pseudomonas fermentation broth with a concentration of 30%-50% is injected from a water injection well, normal water injection is performed for 5-7 days, then the injection is stopped for 1-2 days, normal water injection is performed again for 10-14 days, and one cycle of injection is completed.

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