Enterococcus faecium XL and application thereof

Abstract

The invention discloses an enterococcus faecium XL, and the preservation number of the enterococcus faecium XL is CGMCC (China General Microbiological Culture Collection Center) No. 28133. The enterococcus faecium XL and the application thereof have the following beneficial effects: 1, the enterococcus faecium XL provided by the invention can grow and metabolize under an oil reservoir condition to generate lactic acid organic acid, dissolve carbonate in a rock stratum, increase the permeability of the rock stratum, degrade heavy components in crude oil, relieve crude oil blockage in an oil layer, increase the liquid amount and improve the oil reservoir quality; the oil recovery rate is improved; 2, the enterococcus faecium XL is applied to microorganism single-well throughput, the average single-well oil increasing amount is greater than 400 tons, and the validity period is more than 200 days; and 3, the enterococcus faecium XL is applied to microbial oil displacement, the average daily oil increase of a well group is more than 5 tons, and the water content drop amplitude is more than 5%.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of energy biotechnology and environmental biotechnology, and particularly relates to Enterococcus faecium XL and application thereof. BACKGROUND

[0002] With the continuous development of oil reservoirs, the proportion of domestic medium and low permeability reservoirs is becoming larger and larger. At present, medium and low permeability reservoirs in China have accounted for more than half of the proven reserves, and have become the main battlefield of oil and gas development and construction. Due to the low permeability of the reservoir and the large seepage resistance of heavy oil, it is in the development stage of low oil production rate and low recovery rate.

[0003] During the development of medium and low permeability reservoirs, due to the influence of geology, engineering and drainage factors, the fluid injected into the formation of the injection well is not compatible with the formation water, and after mixing, inorganic salt precipitates (such as calcium carbonate) will block the reservoir channel; during oil and gas production, heavy components in crude oil are continuously precipitated, causing damage to the oil layer.

[0004] At present, acidizing and fracturing methods are often used for stimulation.

[0005] For example, Chinese patent CN 101089118B discloses an acidizing agent composition, which contains component A, component B, component C, component D and water, the component A is selected from at least one of HCl, phosphoric acid, HNO3, organic carboxylic acid, the component B is HF or a compound capable of generating HF, the component C is an organic amine phosphonic acid, and the component D is an organic nitrogen-containing compound capable of forming an ammonium salt with acid or an ammonium salt thereof. However, the traditional acidizing plugging removal measure can only act on the near wellbore zone, and it is difficult to penetrate into the formation, especially for low permeability and seriously plugged layers, and it cannot better solve the problem of organic plugging in the reservoir, and it is easy to corrode the casing, which has safety and environmental protection hidden dangers.

[0006] For example, Chinese patent application CN 117005840A discloses a fracturing method for water control and oil increase in low-permeability high-water-saturation reservoirs, which is implemented according to the following steps: step 1, concentrated perforation in the upper part of the reservoir; step 2, nano-sphere solution is used as a pre-treatment fluid and is injected into the reservoir; step 3, pressure measurement is performed after pump stopping and well closing, and after the nano-spheres are fully expanded, the pore throat channels of the high-water-content section of the reservoir are blocked; step 4, fracturing tools are lowered for fracturing operation, and general hydraulic fracturing operation is performed on the perforated layer section. The present application vertically blocks the high-water-content section of the reservoir by means of fixed-point perforation, nano-sphere water plugging material pre-treatment and hydraulic fracturing, reduces water production, and achieves the purpose of water control and oil increase in low-permeability high-water-saturation reservoirs. However, due to the complex contact relationship between the edge and bottom water of the reservoir, it is difficult to control water and stabilize oil by fracturing measures.

[0007] Therefore, how to solve the plugging of the oil layer and improve the formation conductivity, so as to effectively improve the oil recovery rate, has become a problem that needs to be solved urgently.

[0008] Microbial oil recovery is an effective green and environmentally friendly technology for improving oil recovery after water flooding, thermal recovery and chemical flooding. It is a technology for improving oil production or recovery rate by using microorganisms and their metabolites. For example, Chinese invention patent application CN 106047728A discloses a composite microbial profile control agent, its preparation method and application. The composite microbial profile control agent comprises, by mass percentage: 5-10% of microbial profile control agent; 0.2-0.3% of corn particles; 0.2-0.5% of high molecular polymer; 0-0.2% of auxiliary agent; and the balance of water.

[0009] In view of the above problems, breeding of hydrocarbon-utilizing acid-producing microorganisms adapted to anaerobic environment of oil reservoirs can enter the deep anaerobic zone of the oil reservoir, and through reproduction and metabolism, on the one hand, it can degrade heavy components of crude oil covering on the rock, reduce organic blockage, and be beneficial to the production of crude oil in small pores; on the other hand, it can produce organic acid to dissolve carbonate in the rock, increase the permeability of the rock, and provide a new technical option for further improving the recovery rate of medium and low permeability oil reservoirs, which has broad application prospects.

[0010] Enterococcus faecium is widely distributed in nature and is a gram-positive coccus. It can regulate the balance of intestinal flora, promote the body to produce antibacterial active substances, and kill pathogenic bacteria. It can also improve the digestibility of nutrients, enhance the absorption of nutrients by animals, and has immunomodulatory effects.

[0011] At present, the uses of Enterococcus faecium mainly focus on fermented feed, kitchen waste degradation and water pollution treatment. For example, Chinese invention patent application CN 102093967A discloses the application of a strain of Enterococcus faecium in carbohydrate fermentation and cholesterol degradation, which can increase the food intake of mink and enhance the immune function of the body. For another example, Chinese invention patent application CN 112322530A discloses a functional bacterial agent for degrading seafood kitchen waste, and its preparation method and application. The functional bacteria are used for synergistic fermentation to degrade macromolecular components (such as proteins and fats) in seafood waste into small molecular active substances. The fermented and dried organic fertilizer promotes the growth of crops and turns waste into treasure. For another example, Chinese invention patent application CN 112322539A discloses a strain of Enterococcus faecium screened from sea mud, which has strong effects of degrading nitrite and ammonia nitrogen, and the degradation effect is stable and continuous, which has good application prospects in water treatment.

[0012] In summary, the uses of Enterococcus faecium mainly focus on fermented feed, kitchen waste degradation and water pollution treatment, and there is no report on its application in oil exploitation. SUMMARY

[0013] Purpose of the invention: In view of the deficiencies of the above-mentioned prior art, one of the purposes of the present invention is to provide an anaerobic, hydrocarbon-acid-producing Enterococcus faecium XL. The anaerobic, hydrocarbon-acid-producing Enterococcus faecium XL is a new strain that can reproduce and metabolize under anaerobic conditions, can grow in situ under reservoir conditions, degrade crude oil, and produce organic acids, thereby improving rock properties and increasing crude oil recovery.

[0014] A second object of the present invention is to provide a bacterial agent containing the above-mentioned microbial strain or a fermentation broth prepared by the microbial strain.

[0015] The third purpose of the present invention is to apply the above-mentioned microbial strain, its bacterial agent or its fermentation liquid to the microbial single-well throughput treatment of oil wells in medium- and low-permeability water-driven oil reservoirs. The microorganisms degrade crude oil and improve crude oil fluidity. The organic acid produced can remove carbonate blockage deep in the formation, thereby achieving the purpose of increasing single-well productivity.

[0016] The fourth purpose of the present invention is to apply the above-mentioned microbial strain, its bacterial agent or its fermentation liquid to microbial oil recovery in medium and low permeability reservoirs, reproduce and metabolize the heavy components of crude oil, and at the same time, organic acids dissolve part of the carbonates in the rock formation, increase the permeability of the rock formation, and improve the recovery rate of medium and low permeability reservoirs.

[0017] Technical solution: A strain of Enterococcus faecium XL, whose preservation number is CGMCC No.28133.

[0018] The detailed deposit information of a strain of Enterococcus faecium XL is as follows:

[0019] The microbial deposit number is: CGMCC NO.28133;

[0020] The preservation time is: August 9, 2023;

[0021] Classification name: Enterococcus faecium;

[0022] Depository: General Microbiology Center, China Microbiology Collection Administration;

[0023] Deposit address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China.

[0024] A bacterial agent, the active ingredient of which is the above-mentioned Enterococcus faecium.

[0025] A fermentation liquid is a liquid obtained by metabolizing the above-mentioned Enterococcus faecium in a fermentation medium, wherein the active ingredient of the fermentation liquid is the above-mentioned Enterococcus faecium.

[0026] Further, the fermentation medium comprises: a carbon source of at least 10 g / L, a nitrogen source of at least 2 g / L, a phosphorus source of at least 0.2 g / L, an inorganic salt of at least 0.2 g / L, a pH value of 6.5-7.5, and the balance being water, and / or

[0027] The carbon source is at least one of sucrose, glucose, dextrin, molasses, galactose, arabinose, melibiose, xylose, and maltose, and / or

[0028] The nitrogen source is at least one of corn steep liquor, soybean meal, peptone, beef extract, yeast extract, urea, ammonium chloride, and / or

[0029] The phosphorus source is at least one of dipotassium hydrogen phosphate, diammonium hydrogen phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, ammonium dihydrogen phosphate, and / or

[0030] The inorganic salt is at least one of magnesium sulfate, sodium acetate, potassium sulfate, potassium chloride, sodium chloride, and magnesium chloride.

[0031] Further, the fermentation conditions of Enterococcus faecium are: an inoculation amount of at least 0.2% V / V, an inoculation age of at least 5 h, a fermentation temperature of 35-45°C, and static culture for at least 12 h.

[0032] Preferably, the fermentation conditions of Enterococcus faecium are: an inoculation amount of 1-5% V / V, an inoculation age of 10-15 h, a fermentation temperature of 35-45°C, and static culture for 24-48 h.

[0033] The application of substance I and / or substance II and / or substance III in oil exploitation, wherein:

[0034] The substance I is the Enterococcus faecium described above;

[0035] The substance II is the bacterial agent described above;

[0036] The substance III is the fermentation broth described above.

[0037] The application of substance I and / or substance II and / or substance III in microbial oil production of oil wells in low-to-medium permeability water-drive reservoirs, wherein:

[0038] The substance I is the Enterococcus faecium described above;

[0039] The substance II is the bacterial agent described above;

[0040] The substance III is the fermentation broth described above.

[0041] The application of substance I and / or substance II and / or substance III in microbial single-well huff and puff in low-to-medium permeability water-drive reservoirs, wherein:

[0042] The substance I is the Enterococcus faecium described above;

[0043] Substance II is the above-mentioned bacterial agent;

[0044] Substance III is the above-mentioned fermentation liquor.

[0045] Further, the above-mentioned application comprises the following steps:

[0046] (1) Calculate the total injection volume V of Enterococcus faecium fermentation liquor, fermentation medium and water, whose calculation formula is as follows:

[0047] V = 3.14R 2 Hфβ, wherein:

[0048] V - total injection volume of Enterococcus faecium fermentation liquor, fermentation medium and water, unit: m 3

[0049] R - treatment radius, unit: m, value range: 5-10;

[0050] H - effective thickness, unit: m;

[0051] ф - porosity, dimensionless;

[0052] β - dosage coefficient, value range: 0.6-1.2;

[0053] (2) Respectively take appropriate amount of Enterococcus faecium fermentation liquor, fermentation medium and water, mix uniformly to obtain a mixed liquor, then inject the mixed liquor into the formation through the oil casing annulus of the oil well, and then inject formation water for at least 20m 3 , preferably 50-100m 3 , injection speed is at least 2m 3 / h, preferably 5-10m 3 / h, shut-in culture for at least 3 days, preferably 5-20d, then open well production, wherein:

[0054] The total volume of the mixed liquor is the total injection volume V calculated in step (1);

[0055] V 屎肠球菌发酵液 :V 发酵培养基 = 1:1-20;

[0056] The volume concentration of Enterococcus faecium fermentation liquor in the mixed liquor is 3%-30%.

[0057] The organic acid produced by the growth and metabolism of Enterococcus faecium is used to play a function of plugging removal on the oil well, and finally improve the single well productivity.

[0058] Further, the above-mentioned application comprises the following steps:

[0059] (1) Calculate the total injection volume V of Enterococcus faecium fermentation liquor, fermentation medium and water, whose calculation formula is as follows: ​

[0060] V = 3.14R 2 H = Hf + Hb, wherein:

[0061] V - total volume of injection of Enterococcus faecalis fermentation liquor, fermentation medium and water, in m 3 ;

[0062] R - treatment radius, in m, in the range of 5-10;

[0063] H - effective thickness, in m;

[0064] f - porosity, dimensionless;

[0065] b - dosage coefficient, in the range of 0.6-1.2;

[0066] (2) First slug: appropriate amounts of Enterococcus faecalis fermentation liquor, fermentation medium and water are weighed and mixed to obtain a mixed liquor, which is injected into the formation through the oil well casing annulus, and then formation water is injected for at least 20 m 3 , preferably 50-100 m 3 , at an injection rate of at least 2 m 3 / h, preferably 5-10 m 3 / h, and the well is shut in for at least 3 d, preferably 10-20 d, wherein:

[0067] the total volume of the mixed liquor is the total injection volume V calculated in step (1);

[0068] V 屎肠球菌发酵液 :V 发酵培养基 = 1:1-20;

[0069] the volume concentration of Enterococcus faecalis fermentation liquor in the mixed liquor is 3%-30%;

[0070] (3) Inject a second slug: first inject water for at least 10 m 3 , preferably 20-50 m 3 , then mix appropriate amounts of bio-emulsifier fermentation liquor, activator and water to obtain a mixed liquor, which is injected into the bottom layer through the oil well casing annulus, and then formation water is injected for at least 20 m 3 , preferably 80-150 m 3 , at an injection rate of at least 2 m 3 / h, preferably 10-30 m 3 / h, and the well is shut in for at least 5 d, preferably 10-15 d, and then the well is opened for production, wherein:

[0071] the volume of the mixed liquor is at least 100 m 3 , preferably 100-300 m 3 ;

[0072] The volume concentration of the bio-emulsifier fermentation liquor in the mixed solution is 5% to 20%;

[0073] V 生物乳化剂发酵液 :V 激活剂 =1:1 to 10;

[0074] The bio-emulsification fermentation liquor is one of Rhodococcus fermentation liquor, Arthrobacter fermentation liquor, Bacillus tekriensis fermentation liquor, and Bacillus pallidus fermentation liquor.

[0075] The activator comprises at least 5 g / L of carbon source, at least 2 g / L of nitrogen source, at least 0.1 g / L of phosphorus source, at least 0.05 g / L of growth factor, pH value of 6.0 to 8.0, and the rest is water, wherein:

[0076] The carbon source is at least one of glucose, sucrose, starch, cassava powder, secondary powder, glycerol, and fructose, and / or

[0077] The nitrogen source is at least one of protein peptone, sodium nitrate, corn syrup, bean cake powder, beef extract, yeast powder, urea, ammonium chloride, ammonia water, and ammonium sulfate, and / or

[0078] The phosphorus source is at least one of disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, diammonium hydrogen phosphate, sodium dihydrogen phosphate, and ammonium dihydrogen phosphate, and / or

[0079] The growth factor is at least one of vitamin, small molecule peptide, amino acid, aromatic hydrocarbon, alkane, and xylitol.

[0080] Rhodococcus, Arthrobacter, Bacillus tekriensis, and Bacillus pallidus can produce bio-emulsifiers, thereby reducing the viscosity of crude oil.

[0081] The organic acid produced by the growth and metabolism of Enterococcus faecalis XL plays a function of plugging removal on an oil well, and the bio-surfactant produced by the growth and metabolism of the bio-emulsification bacteria realizes the emulsification and oil washing of crude oil, thereby improving the single well productivity, and is particularly suitable for thick oil low-efficiency wells with underground crude oil viscosity of ≥100 mPa·s.

[0082] The application of substance I and / or substance II and / or substance III in microbial oil displacement in a low-permeability water-drive oil reservoir, wherein:

[0083] The substance I is the Enterococcus faecalis described above.

[0084] The substance II is the bacterial agent described above.

[0085] The substance III is the fermentation liquor described above.

[0086] Further, the application comprises the following steps:

[0087] Appropriate amounts of Enterococcus faecium fermentation liquid, fermentation medium and water are weighed respectively, mixed evenly to obtain a mixed solution, and then the mixed solution is injected into the water injection well, and then normal water injection is performed for at least 3 days, preferably 5 to 10 days, to push the Enterococcus faecium into the deep part of the oil reservoir, and the injection is stopped and cultured for at least 5 days, preferably 8 to 20 days, to allow the Enterococcus faecium to fully reproduce and metabolize in the oil reservoir, and then normal water injection is performed for at least 7 days, preferably 10 to 15 days, to complete one injection cycle, wherein:

[0088] The total injection volume of Enterococcus faecium fermentation liquid, fermentation medium and water is at least 200m 3 , preferably 500 to 1000 m 3 ;

[0089] V 屎肠球菌发酵液 :V 发酵培养基 =1:1~30;

[0090] The volume concentration of the Enterococcus faecium fermentation liquid in the mixed liquid is 5% to 50%.

[0091] Furthermore, the specific steps of the application include:

[0092] First plug: Weigh appropriate amounts of Enterococcus faecium fermentation liquid, fermentation medium and water, mix them evenly to obtain a mixed liquid, and then inject the mixed liquid into the injection well, wherein:

[0093] The total injection volume of Enterococcus faecium fermentation liquid, fermentation medium and water is at least 200m 3 , preferably 500 to 1000 m 3 ;

[0094] V 屎肠球菌发酵液 :V 发酵培养基 =1:1~30;

[0095] The volume concentration of the Enterococcus faecium fermentation liquid in the mixed liquid is 5% to 50%;

[0096] Then inject the second plug: first inject water for at least 1 day, preferably 3 to 5 days, then weigh appropriate amounts of bio-emulsifier fermentation liquid, activator and water, mix them evenly to obtain a mixed solution, inject the mixed solution into the injection well, then inject water normally for at least 3 days, preferably 8 to 15 days, stop injection and cultivate for at least 7 days, preferably 10 to 25 days, and then inject water normally for at least 7 days, preferably 15 to 20 days, to complete one cycle of injection, wherein:

[0097] V 生物乳化剂发酵液 :V 激活剂 =1:2~5;

[0098] The total injection volume of bio-emulsifier fermentation liquid, activator and water is at least 100m 3 , preferably 300~800m 3 ;

[0099] The volume concentration of the bioemulsifier fermentation liquor in the mixed solution is 5% to 30%;

[0100] The bioemulsifier fermentation liquor is one of Rhodococcus fermentation liquor, Arthrobacter fermentation liquor, Bacillus tekriensis fermentation liquor and Bacillus pallidus fermentation liquor;

[0101] The activator comprises: a carbon source of at least 5 g / L, a nitrogen source of at least 2 g / L, a phosphorus source of at least 0.1 g / L, a growth factor of at least 0.05 g / L, a pH value of 6.0 to 8.0, and the balance is water, wherein:

[0102] The carbon source is at least one of glucose, sucrose, starch, cassava powder, secondary powder, glycerol and fructose, and / or

[0103] The nitrogen source is at least one of peptone, sodium nitrate, corn syrup, bean cake powder, beef extract, yeast powder, urea, ammonium chloride, ammonia water and ammonium sulfate, and / or

[0104] The phosphorus source is at least one of disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, diammonium hydrogen phosphate, sodium dihydrogen phosphate and ammonium dihydrogen phosphate, and / or

[0105] The growth factor is at least one of vitamins, small molecule peptides, amino acids, aromatic hydrocarbons, alkanes and xylitol.

[0106] Rhodococcus, Arthrobacter, Bacillus tekriensis and Bacillus pallidus can produce bioemulsifiers, thereby reducing the viscosity of crude oil.

[0107] Enterococcus faecalis XL grows and metabolizes to produce organic acids to dissolve and corrode the formation, improve the permeability of the formation, and realize the emulsification and oil washing of crude oil through the bioemulsifier produced by the growth and metabolism of the bioemulsifier bacteria, thereby synergistically improving the production of the well group. It is especially suitable for thick oil well groups with underground crude oil viscosity ≥100 mPa·s.

[0108] A product, the active ingredient of which is substance I and / or substance II and / or substance III;

[0109] Substance I is the Enterococcus faecalis described above;

[0110] Substance II is the microbial agent described above;

[0111] Substance III is the fermentation liquor described above;

[0112] The function of the product is (a) or (b) or (c) or (d):

[0113] (a) oil exploitation;

[0114] (b) microbial oil recovery;

[0115] (c) microbial single well huff and puff;

[0116] (d) microbial oil displacement.

[0117] An oil recovery method, comprising the following steps: adding substance I and / or substance II and / or substance III in the oil production process;

[0118] The substance I is the Enterococcus faecium described above.

[0119] The substance II is the bacterial agent described above.

[0120] The substance III is the fermentation liquor described above.

[0121] Beneficial effects: the Enterococcus faecium XL and the application thereof disclosed by the application have the following beneficial effects:

[0122] 1. The Enterococcus faecium XL provided by the application can grow and metabolize to produce lactic acid organic acid under the oil reservoir condition, dissolve the carbonate in the rock layer, increase the rock layer permeability, can degrade the heavy component of crude oil, can remove the crude oil blockage in the oil layer, improve the liquid volume, and improve the oil recovery rate.

[0123] 2. The Enterococcus faecium XL is applied to microbial single well huff and puff, the average single well oil increase is greater than 400 tons, and the effective period is more than 200 days.

[0124] 3. The Enterococcus faecium XL is applied to microbial oil displacement, the well group average daily oil increase is more than 5 tons, and the water content reduction amplitude is more than 5%. BRIEF DESCRIPTION OF DRAWINGS

[0125] Figure 1 It is the colony morphology diagram of the Enterococcus faecium XL disclosed by the application.

[0126] Figure 2 It is the cell morphology diagram of the Enterococcus faecium XL disclosed by the application.

[0127] Figure 3 It is the phylogenetic tree of the Enterococcus faecium XL disclosed by the application. DETAILED DESCRIPTION

[0128] The specific embodiment of the application is described in detail as follows.

[0129] The ranges disclosed herein are intended to be "open" ranges, unless expressly specified otherwise. For example, a range of "10-50" is intended to be a range of "greater than or equal to 10 and less than or equal to 50," not "greater than or equal to 10, and less than or equal to 50," unless expressly stated otherwise. It is further to be understood that all values and / or ranges between the recited values qualify as options for the variable. For example, a numerical range of "0-5" recited as a range for a variable indicates that "0-5" and all intervening numerical values of this variable, such as "1-4," "1-3," "1-2," "2-5," "3-5," and "4-5," are contemplated for this variable.

[0130] Unless otherwise specified, all embodiments and optional embodiments of the present application can be combined with each other to form new technical solutions.

[0131] Unless otherwise specified, all technical features and optional technical features of the present application can be combined with each other to form new technical solutions.

[0132] Unless otherwise specified, all steps of the present application can be performed in sequence or randomly, and are preferably performed in sequence. For example, the method comprises steps (a) and (b), which means that the method can comprise steps (a) and (b) performed in sequence, or steps (b) and (a) performed in sequence. For example, the method also comprises step (c), which means that step (c) can be added to the method in any order, for example, the method can comprise steps (a), (b) and (c), or steps (a), (c) and (b), or steps (c), (a) and (b), etc.

[0133] Unless otherwise specified, "including" and "containing" mentioned in the present application means open or closed. For example, "including" and "containing" can mean that other components not listed can also be included or contained, or only the listed components can be included or contained.

[0134] Unless otherwise specified, the reaction is carried out at room temperature and normal pressure.

[0135] Unless otherwise specified, all parts or percentages are volume parts or volume percentages.

[0136] In the present application, all the substances used are known substances, which can be purchased or synthesized by known methods.

[0137] In the present application, all the devices or apparatuses used are conventional devices or apparatuses known in the field, which can be purchased.

[0138] In the following specific examples, the fermentation medium used is sucrose 10 g / L, corn syrup 5 g / L, potassium phosphate dibasic 0.5 g / L, diammonium hydrogen citrate 1 g / L, magnesium sulfate 0.1 g / L, sodium acetate 1 g / L, pH 7.0, and the rest is water, unless otherwise specified.

[0139] In the following specific examples, the activator used is glucose 12 g / L, proteose peptone 4 g / L, sodium nitrate 8 g / L, sodium phosphate dibasic 1.2 g / L, potassium phosphate monobasic 0.8 g / L, growth factor 0.4 g / L, pH 7.5, wherein:

[0140] The growth factor is a vitamin.

[0141] In the following specific examples, the volume concentration of the fermentation broth of Enterococcus faecium XL = (volume of the fermentation broth of Enterococcus faecium XL) / (volume of the fermentation broth of Enterococcus faecium XL + volume of the fermentation medium + volume of water) * 100%.

[0142] An Enterococcus faecium XL, with the preservation number of CGMCC No. 28133.

[0143] A microbial agent, wherein the effective component is the Enterococcus faecium described above.

[0144] A fermentation broth, which is a liquid metabolized by the Enterococcus faecium described above in a fermentation medium, wherein the active component of the fermentation broth is the Enterococcus faecium described above.

[0145] Further, the fermentation medium comprises at least 10 g / L of carbon source, at least 2 g / L of nitrogen source, at least 0.2 g / L of phosphorus source, at least 0.2 g / L of inorganic salt, pH 6.5-7.5, and the rest is water, and / or

[0146] The carbon source is at least one of sucrose, glucose, dextrin, molasses, galactose, arabinose, melibiose, xylose, and maltose, and / or

[0147] The nitrogen source is at least one of corn syrup, soybean meal, proteose peptone, beef extract, yeast powder, urea, and ammonium chloride, and / or

[0148] The phosphorus source is at least one of dipotassium hydrogen phosphate, diammonium hydrogen phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, ammonium dihydrogen phosphate, and / or

[0149] The inorganic salt is at least one of magnesium sulfate, sodium acetate, potassium sulfate, potassium chloride, sodium chloride, and magnesium chloride.

[0150] In one embodiment, the fermentation medium comprises: 10 g / L of a carbon source, 2 g / L of a nitrogen source, 0.2 g / L of a phosphorus source, 0.2 g / L of an inorganic salt, a pH of 6.5, and the balance being water, and / or

[0151] The carbon source is sucrose, and / or

[0152] The nitrogen source is corn steep liquor, and / or

[0153] The phosphorus source is dipotassium hydrogen phosphate, and / or

[0154] The inorganic salt is magnesium sulfate.

[0155] In another embodiment, the fermentation medium comprises: 40 g / L of a carbon source, 10 g / L of a nitrogen source, 10 g / L of a phosphorus source, 2 g / L of an inorganic salt, a pH of 7.5, and the balance being water, and / or

[0156] The carbon source is glucose, and / or

[0157] The nitrogen source is soybean meal, and / or

[0158] The phosphorus source is diammonium hydrogen phosphate, and / or

[0159] The inorganic salt is sodium acetate.

[0160] In another embodiment, the fermentation medium comprises: 20 g / L of a carbon source, 5 g / L of a nitrogen source, 5 g / L of a phosphorus source, 1 g / L of an inorganic salt, a pH of 7, and the balance being water, and / or

[0161] The carbon source is dextrin, and / or

[0162] The nitrogen source is peptone, and / or

[0163] The phosphorus source is disodium hydrogen phosphate, and / or

[0164] The inorganic salt is potassium sulfate.

[0165] In another embodiment, the fermentation medium comprises: 10 g / L of a carbon source, 2 g / L of a nitrogen source, 0.2 g / L of a phosphorus source, 0.2 g / L of an inorganic salt, a pH of 7, and the balance being water, and / or

[0166] The carbon source is molasses, and / or

[0167] The nitrogen source is beef extract, and / or

[0168] The phosphorus source is sodium dihydrogen phosphate, and / or

[0169] The inorganic salt is potassium chloride.

[0170] In another embodiment, the fermentation medium comprises: 10 g / L of the carbon source, 2 g / L of the nitrogen source, 0.2 g / L of the phosphorus source, 0.2 g / L of the inorganic salt, pH 7, and the balance being water, and / or

[0171] The carbon source is galactose, and / or

[0172] The nitrogen source is yeast extract, and / or

[0173] The phosphorus source is potassium dihydrogen phosphate, and / or

[0174] The inorganic salt is sodium chloride.

[0175] In another embodiment, the fermentation medium comprises: 20 g / L of the carbon source, 5 g / L of the nitrogen source, 0.8 g / L of the phosphorus source, 0.6 g / L of the inorganic salt, pH 6.5, and the balance being water, and / or

[0176] The carbon source is arabinose, and / or

[0177] The nitrogen source is urea, and / or

[0178] The phosphorus source is ammonium dihydrogen phosphate, and / or

[0179] The inorganic salt is magnesium chloride.

[0180] In another embodiment, the fermentation medium comprises: 10 g / L of the carbon source, 2 g / L of the nitrogen source, 0.2 g / L of the phosphorus source, 0.2 g / L of the inorganic salt, pH 7.5, and the balance being water, and / or

[0181] The carbon source is melibiose, and / or

[0182] The nitrogen source is ammonium chloride, and / or

[0183] The phosphorus source is diammonium hydrogen citrate, and / or

[0184] The inorganic salt is a mixture of magnesium sulfate, sodium acetate, potassium sulfate, potassium chloride, sodium chloride, and magnesium chloride in equal mass ratios.

[0185] In another embodiment, the fermentation medium comprises: a carbon source of 10 g / L, a nitrogen source of 2 g / L, a phosphorus source of 0.2 g / L, an inorganic salt of 0.2 g / L, a pH of 7, and the balance being water, and / or

[0186] The carbon source is a mixture of sucrose, glucose, dextrin, molasses, galactose, arabinose, melibiose, xylose, maltose in equal mass ratio, and in yet another embodiment, the carbon source is xylose. In yet another embodiment, the carbon source is maltose. And / or

[0187] The nitrogen source is a mixture of corn steep liquor, soybean meal, peptone, beef extract, yeast extract, urea, ammonium chloride in equal mass ratio, and / or

[0188] The phosphorus source is a mixture of dipotassium hydrogen phosphate, diammonium hydrogen phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, ammonium dihydrogen phosphate, diammonium hydrogen citrate in equal mass ratio, and / or

[0189] The inorganic salt is a mixture of magnesium sulfate, sodium acetate in equal mass ratio.

[0190] Further, the fermentation conditions of Enterococcus faecium are: inoculum of at least 0.2% V / V, inoculum age of at least 5 h, fermentation temperature of 35-45°C, and static culture for at least 12 h.

[0191] Preferably, the fermentation conditions of Enterococcus faecium are: inoculum of 1-5% V / V, inoculum age of 10-15 h, fermentation temperature of 35-45°C, and static culture for 24-48 h.

[0192] In one embodiment, the fermentation conditions of Enterococcus faecium are: inoculum of 0.2% V / V, inoculum age of 5 h, fermentation temperature of 35°C, and static culture for 12 h.

[0193] In another embodiment, the fermentation conditions of Enterococcus faecium are: inoculum of 1% V / V, inoculum age of 10 h, fermentation temperature of 45°C, and static culture for 36 h.

[0194] In another embodiment, the fermentation conditions of Enterococcus faecium are: inoculum of 1% V / V, inoculum age of 10 h, fermentation temperature of 35°C, and static culture for 48 h.

[0195] In another embodiment, the fermentation conditions of Enterococcus faecium are: inoculum of 5% V / V, inoculum age of 15 h, fermentation temperature of 45°C, and static culture for 24 h.

[0196] In another embodiment, the fermentation conditions of Enterococcus faecium are: inoculum of 3% V / V, inoculum age of 12 h, fermentation temperature of 40°C, and static culture for 36 h.

[0197] Use of the substance I and / or the substance II and / or the substance III in oil exploitation, wherein:

[0198] The substance I is Enterococcus faecium as described above;

[0199] The substance II is the bacterial agent as described above;

[0200] The substance III is the fermentation broth as described above.

[0201] Use of the substance I and / or the substance II and / or the substance III in microbial oil production of oil wells in low-to-moderate permeability water-drive reservoirs, wherein:

[0202] The substance I is Enterococcus faecium as described above;

[0203] The substance II is the bacterial agent as described above;

[0204] The substance III is the fermentation broth as described above.

[0205] Use of the substance I and / or the substance II and / or the substance III in microbial single-well huff and puff of oil wells in low-to-moderate permeability water-drive reservoirs, wherein:

[0206] The substance I is Enterococcus faecium as described above;

[0207] The substance II is the bacterial agent as described above;

[0208] The substance III is the fermentation broth as described above.

[0209] Further, the above use comprises the following steps:

[0210] (1) calculating the total injection volume V of the Enterococcus faecium fermentation broth, the fermentation medium and water, and the calculation formula is as follows:

[0211] V = 3.14R 2 Hфβ, wherein:

[0212] V is the total injection volume of the Enterococcus faecium fermentation broth, the fermentation medium and water, and the unit is m 3 ;

[0213] R is the treatment radius, and the unit is m, and the value range is 5-10;

[0214] H is the effective thickness, and the unit is m;

[0215] ф is the porosity, and is dimensionless;

[0216] β is the dosage coefficient, and the value range is 0.6-1.2;

[0217] (2) respectively taking an appropriate amount of the Enterococcus faecium fermentation broth, the fermentation medium and water, mixing uniformly to obtain a mixed liquid, and then injecting the mixed liquid into the formation through the oil well casing annulus, and then injecting formation water into the formation for at least 20 m 3(1) in one embodiment, the re-injection formation water 20 m 3 (2) in another embodiment, the re-injection formation water 50 m 3 (3) in another embodiment, the re-injection formation water 100 m 3 (4) at an injection rate of at least 2 m 3 / h (in one embodiment, the injection rate is 2 m 3 / h. In another embodiment, the injection rate is 5 m 3 / h. In another embodiment, the injection rate is 10 m 3 / h), the well is shut in for at least 3 days (in one embodiment, the well is shut in for 3 days. In another embodiment, the well is shut in for 5 days, and in another embodiment, the well is shut in for 20 d) before it is opened for production, wherein:

[0218] The total volume of the mixed solution is the total injection volume V calculated in step (1);

[0219] V 屎肠球菌发酵液 :V 发酵培养基 = 1:1~20, such as V 屎肠球菌发酵液 :V 发酵培养基 = 1:1. In another embodiment, V 屎肠球菌发酵液 :V

[0220] 发酵培养基 = 1:20. In another embodiment, V 屎肠球菌发酵液 :V 发酵培养基 = 1:10.

[0221] The volume concentration of the Enterococcus faecalis fermentation broth in the mixed solution is 3%~30%. In another embodiment, the volume concentration of the Enterococcus faecalis fermentation broth in the mixed solution is 3%. In another embodiment, the volume concentration of the Enterococcus faecalis fermentation broth in the mixed solution is 30%. In another embodiment, the volume concentration of the Enterococcus faecalis fermentation broth in the mixed solution is 10%.

[0222] Further, the above application comprises the following steps:

[0223] (1) calculating the total injection volume V of the Enterococcus faecalis fermentation broth, the fermentation medium and water, according to the following formula:

[0224] V = 3.14R 2 Hфβ, wherein:

[0225] V - the total injection volume of the Enterococcus faecalis fermentation broth, the fermentation medium and water, in m 3 ;

[0226] R - the treatment radius, in m, ranging from 5 to 10;

[0227] H - the effective thickness, in m;

[0228] ф - the porosity, dimensionless.

[0229] β - dosage coefficient, value range 0.6-1.2;

[0230] (2), the first plug: respectively take appropriate amount of Enterococcus faecium fermentation broth, fermentation medium and water, mixed evenly to obtain the mixed liquor, then the mixed liquor through the oil well oil jacket annulus injection formation, and then inject formation water at least 20m 3 , preferably 50-100m 3 , injection rate is at least 2m 3 / h, preferably 5-10m 3 / h, shut-in culture at least 3d, preferably 10-20d, wherein:

[0231] The total volume of the mixed liquor is the total injection volume V calculated in step (1);

[0232] V 屎肠球菌发酵液 :V 发酵培养基 =1:1-20;

[0233] The volume concentration of Enterococcus faecium fermentation broth in the mixed liquor is 3%-30%;

[0234] (3), inject the second plug, first inject water at least 10m 3 , preferably 20-50m 3 , then take appropriate amount of bioemulsifier fermentation broth, activator and water, mixed evenly to obtain the mixed liquor, the mixed liquor through the oil well oil jacket annulus injection bottom layer, and then inject formation water at least 20m 3 , preferably 80-150m 3 , injection rate is at least 2m 3 / h, preferably 10-30m 3 / h, shut-in culture at least 5d, preferably 10-15d after opening well production, wherein:

[0235] The volume of the mixed liquor is at least 100m 3 , preferably 100-300m 3 ;

[0236] In the mixed liquor, the volume concentration of bioemulsifier fermentation broth is 5%-20%;

[0237] V 生物乳化剂发酵液 :V 激活剂 =1:1-10;

[0238] Bioemulsifier fermentation broth is one of Rhodococcus fermentation broth, Arthrobacter fermentation broth, Bacillus tekriensis fermentation broth and Bacillus pallidus fermentation broth;

[0239] The activator includes: carbon source at least 5g / L, nitrogen source at least 2g / L, phosphorus source at least 0.1g / L, growth factor at least 0.05g / L, pH value is 6.0-8.0, the balance is water, wherein:

[0240] The carbon source is at least one of glucose, sucrose, starch, cassava powder, powder, glycerol, fructose, and / or

[0241] The nitrogen source is at least one of peptone, sodium nitrate, corn syrup, bean cake powder, beef extract, yeast powder, urea, ammonium chloride, ammonia, ammonium sulfate, and / or

[0242] The phosphorus source is at least one of disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, diammonium hydrogen phosphate, sodium dihydrogen phosphate, ammonium dihydrogen phosphate, and / or

[0243] The growth factor is at least one of vitamin, small molecule peptide, amino acid, aromatic hydrocarbon, alkane, xylitol.

[0244] In one embodiment, the above application comprises the following steps:

[0245] (1), the total amount V of Enterococcus faecium fermentation broth, fermentation medium and water is calculated, and the calculation formula is as follows:

[0246] V=3.14R 2 Hфβ, wherein:

[0247] V-the total amount of Enterococcus faecium fermentation broth, fermentation medium and water, unit m 3 ;

[0248] R-treatment radius, unit m, the value range is 5-10;

[0249] H-effective thickness, unit m;

[0250] f-porosity, dimensionless;

[0251] β-dosage coefficient, the value range is 0.6-1.2;

[0252] (2), the first plug: respectively take appropriate amount of Enterococcus faecium fermentation broth, fermentation medium and water, mix uniformly to obtain mixed solution, then the mixed solution is injected into the formation through oil well oil jacket annulus, and then 20m 3 of formation water is injected, the injection speed is 2m 3 / h, and the well is closed for 3d, wherein:

[0253] The total volume of the mixed solution is the total amount V calculated in step (1);

[0254] V 屎肠球菌发酵液 :V 发酵培养基= 1:10;

[0255] The volume concentration of Enterococcus faecalis fermentation broth in the mixed solution is 10%;

[0256] (3) re-injection of the second slug, first water injection 10m 3 Then take the appropriate amount of biological emulsifier fermentation broth, activator and water, mix evenly to get the mixed solution, the mixed solution is injected into the bottom layer through the oil well oil jacket annulus, and then 20m 3 of formation water is injected at a speed of 2m 3 / h, the oil well is closed for at least 5 days and then opened for production, wherein:

[0257] The volume of the mixed solution is 100m 3 ;

[0258] In the mixed solution, the volume concentration of biological emulsifier fermentation broth is 10%;

[0259] V 生物乳化剂发酵液 :V 激活剂 = 1:5;

[0260] The biological emulsifier fermentation broth is Rhodococcus fermentation broth;

[0261] The activator comprises: 5g / L of carbon source, 2g / L of nitrogen source, 0.1g / L of phosphorus source, 0.05g / L of growth factor, pH value of 7.0, and the balance is water, wherein:

[0262] The carbon source is a mixture of glucose, sucrose, starch, cassava powder, secondary powder, glycerol and fructose in equal mass ratio, and / or

[0263] The nitrogen source is a mixture of peptone, sodium nitrate, corn syrup, bean cake powder, beef extract, yeast powder, urea, ammonium chloride, ammonia, ammonium sulfate in equal mass ratio, and / or

[0264] The phosphorus source is a mixture of disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, diammonium hydrogen phosphate, sodium dihydrogen phosphate, and ammonium dihydrogen phosphate in equal mass ratio, and / or

[0265] The growth factor is at least one of vitamins, small molecule peptides, amino acids, aromatic hydrocarbons, alkanes and xylitol in equal mass ratio.

[0266] In another embodiment, the above application comprises the following steps:

[0267] (1) Calculate the total injection volume V of Enterococcus faecalis fermentation broth, fermentation medium and water, the calculation formula is as follows:

[0268] V = 3.14R 2 Hфβ, wherein:

[0269] V—total volume of the fecal enterococcus fermentation liquor, the fermentation medium and water, unit: m 3 ;

[0270] R—treatment radius, unit: m, value range: 5-10;

[0271] H—effective thickness, unit: m;

[0272] φ—porosity, dimensionless;

[0273] β—use coefficient, value range: 0.6-1.2;

[0274] (2), the first slug: respectively take an appropriate amount of fecal enterococcus fermentation liquor, fermentation medium and water, mix uniformly to obtain a mixed liquor, then the mixed liquor is injected into the formation through the oil well oil jacket annulus, and then 50m 3 of formation water is injected at a speed of 5m 3 / h, and the well is closed for 10d, wherein:

[0275] the total volume of the mixed liquor is the total injection volume V calculated in step (1);

[0276] V 屎肠球菌发酵液 :V 发酵培养基 =1:1;

[0277] The volume concentration of the fecal enterococcus fermentation liquor in the mixed liquor is 3%;

[0278] (3), the second slug is injected again, first 20m 3 of water is injected, then an appropriate amount of biological emulsifier fermentation liquor, activator and water are mixed uniformly to obtain a mixed liquor, the mixed liquor is injected into the bottom layer through the oil well oil jacket annulus, and then 80m 3 of formation water is injected at a speed of 10m 3 / h, and the well is closed for 10d after the well is opened for production, wherein:

[0279] the volume of the mixed liquor is 100m 3 ;

[0280] In the mixed liquor, the volume concentration of the biological emulsifier fermentation liquor is 5%;

[0281] V 生物乳化剂发酵液 :V 激活剂 =1:1;

[0282] The biological emulsifier fermentation liquor is arthrobacter fermentation liquor;

[0283] The activator comprises: 10g / L of carbon source, 6g / L of nitrogen source, 10g / L of phosphorus source, 0.1g / L of growth factor, pH value of 6.0, and the balance is water, wherein:

[0284] the carbon source is glucose, and / or

[0285] the nitrogen source is peptone, and / or

[0286] the phosphorus source is disodium hydrogen phosphate, and / or

[0287] the growth factor is vitamin.

[0288] In another embodiment, the above application comprises the following steps:

[0289] (1) calculating the total volume V of injection of Enterococcus faecalis fermentation liquor, fermentation medium and water, the calculation formula is as follows:

[0290] V = 3.14R 2 Hфβ, wherein:

[0291] V - the total volume of injection of Enterococcus faecalis fermentation liquor, fermentation medium and water, in m 3 ;

[0292] R - treatment radius, in m, the value range is 5-10;

[0293] H - effective thickness, in m;

[0294] f - porosity, dimensionless;

[0295] β - dosage coefficient, the value range is 0.6-1.2;

[0296] (2) first slug: respectively take an appropriate amount of Enterococcus faecalis fermentation liquor, fermentation medium and water, mix uniformly to obtain a mixed liquor, then inject the mixed liquor into the stratum through the oil casing annulus of the oil well, and then inject 100 m 3 of formation water, the injection speed is 10 m 3 / h, shut-in culture for 20 days, wherein:

[0297] the total volume of the mixed liquor is the total volume of injection V calculated in step (1);

[0298] V 屎肠球菌发酵液 :V 发酵培养基 = 1:20;

[0299] the volume concentration of Enterococcus faecalis fermentation liquor in the mixed liquor is 30%;

[0300] (3) inject a second slug, first inject 50 m 3 of water, then take an appropriate amount of bio-emulsifier fermentation liquor, activator and water, mix uniformly to obtain a mixed liquor, inject the mixed liquor into the bottom layer through the oil casing annulus of the oil well, and then inject 150 m 3 of formation water, the injection speed is 30 m 3 / h. / h, the well is opened to produce after 15 days of shut-in culture, wherein:

[0301] The volume of the mixed solution is 300m 3 ;

[0302] The volume concentration of the bio-emulsifier fermentation liquor in the mixed solution is 20%;

[0303] V 生物乳化剂发酵液 :V 激活剂 =1:10;

[0304] The bio-emulsifier fermentation liquor is Bacillus tekronensis fermentation liquor;

[0305] The activator comprises: a carbon source of 40g / L, a nitrogen source of 10g / L, a phosphorus source of 10g / L, a growth factor of at least 0.2g / L, a pH value of 8.0, and the balance is water, wherein:

[0306] The carbon source is sucrose, and / or

[0307] The nitrogen source is sodium nitrate, and / or

[0308] The phosphorus source is potassium dihydrogen phosphate, and / or

[0309] The growth factor is a small molecule peptide.

[0310] In another embodiment, the above application comprises the following steps:

[0311] (1) Calculate the total injection amount V of Enterococcus faecium fermentation liquor, fermentation medium and water, and the calculation formula is as follows:

[0312] V=3.14R 2 Hфβ, wherein:

[0313] V-the total injection amount of Enterococcus faecium fermentation liquor, fermentation medium and water, unit: m 3 ;

[0314] R-treatment radius, unit: m, value range: 5-10;

[0315] H-effective thickness, unit: m;

[0316] ф-porosity, dimensionless;

[0317] β-dosage coefficient, value range: 0.6-1.2;

[0318] (2) First slug: appropriate amounts of Enterococcus faecium fermentation liquor, fermentation medium and water are weighed respectively, mixed uniformly to obtain a mixed solution, and then the mixed solution is injected into the formation through the oil jacket annulus of the oil well, and then 80m 3 of formation water is injected at an injection speed of 8m3 / h, shut-in for 15 days, wherein:

[0319] The total volume of the mixed solution is the total injection volume V calculated in step (1);

[0320] V 屎肠球菌发酵液 :V 发酵培养基 = 1:10;

[0321] The volume concentration of the Enterococcus faecium fermentation broth in the mixed solution is 10%;

[0322] (3) injecting a second slug, first injecting water 30m 3 Then, mixing an appropriate amount of bio-emulsifier fermentation broth, activator and water to obtain a mixed solution, injecting the mixed solution into the bottom layer through the oil well oil casing annulus, and then injecting 100m 3 of formation water at an injection rate of 20m 3 / h, shut-in for 12 days, and then opening the well for production, wherein:

[0323] The volume of the mixed solution is 200m 3 ;

[0324] In the mixed solution, the volume concentration of the bio-emulsifier fermentation broth is 10%;

[0325] V 生物乳化剂发酵液 :V 激活剂 = 1:5;

[0326] The bio-emulsifier fermentation broth is Bacillus pallidus fermentation broth;

[0327] The activator comprises: a carbon source of 20g / L, a nitrogen source of 20g / L, a phosphorus source of 5g / L, a growth factor of at least 0.2g / L, a pH value of 7.0, and the balance being water, wherein:

[0328] The carbon source is starch; in another embodiment, the carbon source is cassava powder; in another embodiment, the carbon source is wheat bran; in another embodiment, the carbon source is glycerol; in another embodiment, the carbon source is fructose, and / or

[0329] The nitrogen source is corn steep liquor; in another embodiment, the nitrogen source is soybean meal; in another embodiment, the nitrogen source is beef extract; in another embodiment, the nitrogen source is yeast extract; in another embodiment, the nitrogen source is urea; in another embodiment, the nitrogen source is ammonium chloride; in another embodiment, the nitrogen source is ammonia water; in another embodiment, the nitrogen source is ammonium sulfate;

[0330] The phosphorus source is dipotassium hydrogen phosphate; in another embodiment, the phosphorus source is diammonium hydrogen phosphate; in another embodiment, the phosphorus source is sodium dihydrogen phosphate; in another embodiment, the phosphorus source is ammonium dihydrogen phosphate, and / or

[0331] The growth factor is an amino acid; in another embodiment, the growth factor is an aromatic hydrocarbon; in another embodiment, the growth factor is an alkane; in another embodiment, the growth factor is xylitol.

[0332] The application of substance I and / or substance II and / or substance III in microbial oil displacement in low-to-moderate permeability water-drive reservoirs, wherein:

[0333] The substance I is Enterococcus faecium as described above;

[0334] The substance II is the bacterial agent as described above;

[0335] The substance III is the fermentation broth as described above.

[0336] Further, the application comprises the following steps:

[0337] An appropriate amount of Enterococcus faecium fermentation broth, fermentation medium and water are weighed respectively, mixed uniformly to obtain a mixed solution, and then the mixed solution is injected into a water injection well, and then normal water injection is performed for at least 3 days (such as 3 days, 5 days or 10 days), so as to push the Enterococcus faecium into the deep part of the oil reservoir, and then the injection is stopped for at least 5 days (such as 5 days, 8 days, 10 days or 20 days) to allow the Enterococcus faecium to fully reproduce and metabolize in the oil reservoir, and then normal water injection is performed for at least 7 days (such as 7 days, 8 days, 10 days or 15 days) to complete one cycle of injection, wherein:

[0338] The total injection amount of the Enterococcus faecium fermentation broth, the fermentation medium and the water is at least 200 m 3 (For example, 200 m 3 , 500, 1000 m 3 );

[0339] V 屎肠球菌发酵液 :V 发酵培养基 =1:1~30 (such as V 屎肠球菌发酵液 :V 发酵培养基 =1:1 or V 屎肠球菌发酵液 :V 发酵培

[0340] 养基 =1:30 or V 屎肠球菌发酵液 :V 发酵培养基 =1:10);

[0341] The volume concentration of the Enterococcus faecium fermentation broth in the mixed solution is 5%~50% (such as 5% or 50% or 25%).

[0342] Further, the specific steps of the application include:

[0343] The first plug: take appropriate amount of Enterococcus faecium fermentation liquor, fermentation medium and water respectively, mix them to get a mixed liquor, then inject the mixed liquor into the injection well, wherein:

[0344] The total injection amount of Enterococcus faecium fermentation liquor, fermentation medium and water is at least 200m 3 , preferably 500-1000m 3 ;

[0345] V 屎肠球菌发酵液 :V 发酵培养基 =1:1-30;

[0346] The volume concentration of Enterococcus faecium fermentation liquor in the mixed liquor is 5%-50%;

[0347] The second plug: first inject water for at least 1d, preferably 3-5d, then take appropriate amount of bio-emulsifier fermentation liquor, activator and water respectively, mix them to get a mixed liquor, inject the mixed liquor into the injection well, then normally inject water for at least 3d, preferably 8-15d, stop injection for at least 7d, preferably 10-25d, then normally inject water for at least 7d, preferably 15-20d, complete a cycle of injection, wherein:

[0348] V 生物乳化剂发酵液 :V 激活剂 =1:2-5;

[0349] The total injection amount of bio-emulsifier fermentation liquor, activator and water is at least 100m 3 , preferably 300-800m 3 ;

[0350] The volume concentration of bio-emulsifier fermentation liquor in the mixed liquor is 5%-30%;

[0351] The bio-emulsifier fermentation liquor is one of Rhodococcus fermentation liquor, Arthrobacter fermentation liquor, Bacillus tekriensis fermentation liquor and Bacillus pallidus fermentation liquor;

[0352] The activator includes: carbon source at least 5g / L, nitrogen source at least 2g / L, phosphorus source at least 0.1g / L, growth factor at least 0.05g / L, pH value 6.0-8.0, and the rest is water, wherein:

[0353] The carbon source is at least one of glucose, sucrose, starch, cassava powder, secondary powder, glycerol and fructose, and / or

[0354] The nitrogen source is at least one of protein peptone, sodium nitrate, corn syrup, bean cake powder, beef extract, yeast powder, urea, ammonium chloride, ammonia water and ammonium sulfate, and / or

[0355] The phosphorus source is at least one of disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, diammonium hydrogen phosphate, sodium dihydrogen phosphate, and ammonium dihydrogen phosphate, and / or

[0356] The growth factor is at least one of a vitamin, a small molecule peptide, an amino acid, an aromatic hydrocarbon, an alkane, and xylitol.

[0357] In one embodiment, the specific steps of the application include:

[0358] The first plug: an appropriate amount of Enterococcus faecium fermentation liquor, fermentation medium and water are weighed respectively, mixed uniformly to obtain a mixed liquor, and then the mixed liquor is injected into a water injection well, wherein:

[0359] The total injection amount of the Enterococcus faecium fermentation liquor, the fermentation medium and water is 200m 3 ;

[0360] V 屎肠球菌发酵液 :V 发酵培养基 =1:10;

[0361] The volume concentration of the Enterococcus faecium fermentation liquor in the mixed liquor is 10%;

[0362] The second plug is injected: first, water is injected for 1d, then an appropriate amount of bio-emulsifier fermentation liquor, activator and water are weighed respectively, mixed uniformly to obtain a mixed liquor, the mixed liquor is injected into the water injection well, then normal water injection is performed for 3d, injection is stopped for 7d of culture, and normal water injection is performed for at least 7d to complete a cycle of injection, wherein:

[0363] V 生物乳化剂发酵液 :V 激活剂 =1:4;

[0364] The total injection amount of the bio-emulsifier fermentation liquor, the activator and water is 100m 3 ;

[0365] The volume concentration of the bio-emulsifier fermentation liquor in the mixed liquor is 5%;

[0366] The bio-emulsifier fermentation liquor is Rhodococcus fermentation liquor;

[0367] The activator includes: a carbon source of 5g / L, a nitrogen source of 2g / L, a phosphorus source of 0.1g / L, a growth factor of 0.05g / L, a pH value of 7.0, and the balance is water, wherein:

[0368] The carbon source is a mixture of glucose, sucrose, starch, cassava powder, flour, glycerol and fructose in equal mass ratio, and / or

[0369] The nitrogen source is a mixture of equal mass ratio of peptone, sodium nitrate, corn steep liquor, soybean meal, beef extract, yeast extract, urea, ammonium chloride, ammonia, ammonium sulfate, and / or

[0370] The phosphorus source is a mixture of equal mass ratio of disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, diammonium hydrogen phosphate, sodium dihydrogen phosphate, ammonium dihydrogen phosphate, and / or

[0371] The growth factor is a mixture of equal mass ratio of vitamins, small molecule peptides, amino acids, aromatic hydrocarbons, alkanes, and xylitol.

[0372] In another embodiment, the specific steps of the application include:

[0373] The first plug: an appropriate amount of Enterococcus faecium fermentation broth, fermentation medium and water are weighed respectively, mixed uniformly to obtain a mixed solution, and then the mixed solution is injected into the injection well, wherein:

[0374] The total injection amount of Enterococcus faecium fermentation broth, fermentation medium and water is 500;

[0375] V 屎肠球菌发酵液 :V 发酵培养基 =1:1;

[0376] The volume concentration of Enterococcus faecium fermentation broth in the mixed solution is 5%;

[0377] The second plug is injected: first, water is injected for 3d, then an appropriate amount of bio-emulsifier fermentation broth, activator and water are weighed respectively, mixed uniformly to obtain a mixed solution, the mixed solution is injected into the injection well, then normal water injection is carried out for 8d, injection is stopped for 10d of culture, and then normal water injection is carried out for 15d, completing a cycle of injection, wherein:

[0378] V 生物乳化剂发酵液 :V 激活剂 =1:2;

[0379] The total injection amount of bio-emulsifier fermentation broth, activator and water is 300m 3 ;

[0380] The volume concentration of bio-emulsifier fermentation broth in the mixed solution is 5%;

[0381] The bio-emulsifier fermentation broth is Arthrobacter fermentation broth;

[0382] The activator includes: a carbon source of 40g / L, a nitrogen source of 10g / L, a phosphorus source of 10g / L, a growth factor of 2g / L, a pH value of 6.0, and the balance is water, wherein:

[0383] The carbon source is glucose, and / or

[0384] The nitrogen source is peptone, and / or

[0385] The phosphorus source is sodium hydrogen phosphate, and / or

[0386] The growth factor is a vitamin.

[0387] In another embodiment, the specific steps of the application include:

[0388] The first plug: the appropriate amount of Enterococcus faecium fermentation broth, fermentation medium and water are weighed respectively, mixed uniformly to obtain a mixed solution, and then the mixed solution is injected into the injection well, wherein:

[0389] The total injection amount of Enterococcus faecium fermentation broth, fermentation medium and water is 1000m 3 ;

[0390] V 屎肠球菌发酵液 :V 发酵培养基 =1:30;

[0391] The volume concentration of Enterococcus faecium fermentation broth in the mixed solution is 50%;

[0392] The second plug is injected: first, water is injected for 5d, then the appropriate amount of biological emulsifier fermentation broth, activator and water are weighed respectively, mixed uniformly to obtain a mixed solution, the mixed solution is injected into the injection well, then normal water injection is carried out for 15d, injection is stopped for 25d, and then normal water injection is carried out for 20d, to complete a cycle of injection, wherein:

[0393] V 生物乳化剂发酵液 :V 激活剂 =1:5;

[0394] The total injection amount of biological emulsifier fermentation broth, activator and water is 800m 3 ;

[0395] The volume concentration of biological emulsifier fermentation broth in the mixed solution is 30%;

[0396] The biological emulsifier fermentation broth is Bacillus tequilensis fermentation broth;

[0397] The activator includes: a carbon source of 20g / L, a nitrogen source of 6g / L, a phosphorus source of 6g / L, a growth factor of 0.1g / L, a pH value of 8.0, and the balance is water, wherein:

[0398] The carbon source is sucrose, and / or

[0399] The nitrogen source is sodium nitrate, and / or

[0400] The phosphorus source is potassium dihydrogen phosphate, and / or

[0401] The growth factor is a small molecule peptide.

[0402] In another embodiment, the specific steps of the application include:

[0403] The first plug: the appropriate amount of Enterococcus faecium fermentation broth, fermentation medium and water are weighed respectively, mixed uniformly to obtain a mixed solution, and then the mixed solution is injected into the injection well, wherein:

[0404] The total injection amount of Enterococcus faecium fermentation broth, fermentation medium and water is 600m 3 ;

[0405] V 屎肠球菌发酵液 :V 发酵培养基 =1:20;

[0406] The volume concentration of Enterococcus faecium fermentation broth in the mixed solution is 10%;

[0407] The second plug is injected: first, water is injected for 4d, then the appropriate amount of biological emulsifier fermentation broth, activator and water are weighed respectively, mixed uniformly to obtain a mixed solution, the mixed solution is injected into the injection well, then normal water injection is carried out for 10d, injection is stopped for 20d, and then normal water injection is carried out for 18d, completing a cycle of injection, wherein:

[0408] V 生物乳化剂发酵液 :V 激活剂 =1:3;

[0409] The total injection amount of biological emulsifier fermentation broth, activator and water is 600m 3 ;

[0410] The volume concentration of biological emulsifier fermentation broth in the mixed solution is 20%;

[0411] The biological emulsifier fermentation broth is Bacillus pallidus fermentation broth;

[0412] The activator includes: 10g / L of carbon source, 2g / L of nitrogen source, 0.8g / L of phosphorus source, 0.08g / L of growth factor, pH value of 7.0, and the balance is water, wherein:

[0413] The carbon source is starch; in another embodiment, the carbon source is cassava powder; in another embodiment, the carbon source is secondary powder; in another embodiment, the carbon source is glycerol; in another embodiment, the carbon source is fructose, and / or

[0414] The nitrogen source is corn steep liquor; in another embodiment, the nitrogen source is soybean meal; in another embodiment, the nitrogen source is beef extract; in another embodiment, the nitrogen source is yeast extract; in another embodiment, the nitrogen source is urea; in another embodiment, the nitrogen source is aqueous ammonia; in another embodiment, the nitrogen source is ammonium sulfate, in another embodiment, the nitrogen source is ammonium chloride, and / or

[0415] The phosphorus source is dipotassium hydrogen phosphate; in another embodiment, the phosphorus source is diammonium hydrogen phosphate; in another embodiment, the phosphorus source is sodium dihydrogen phosphate; in another embodiment, the phosphorus source is ammonium dihydrogen phosphate, and / or

[0416] The growth factor is an amino acid; in another embodiment, the growth factor is an aromatic hydrocarbon; in another embodiment, the growth factor is an alkane; in another embodiment, the growth factor is xylitol.

[0417] A product, whose active ingredient is substance I and / or substance II and / or substance III;

[0418] Substance I is Enterococcus faecium as described above;

[0419] Substance II is the bacterial agent as described above;

[0420] Substance III is the fermentation broth as described above;

[0421] The function of the product is (a) or (b) or (c) or (d):

[0422] (a) oil production;

[0423] (b) microbial enhanced oil recovery;

[0424] (c) microbial single-well huff and puff;

[0425] (d) microbial flooding.

[0426] An oil production method, comprising the following steps: during the oil production process, substance I and / or substance II and / or substance III are added;

[0427] Substance I is Enterococcus faecium as described above;

[0428] Substance II is the bacterial agent as described above;

[0429] Substance III is the fermentation broth as described above.

[0430] Example 1: Obtainment and identification of Enterococcus faecium XL

[0431] I. Obtainment of the strain

[0432] A strain was isolated from the oil well production fluid sample of Shengli Oilfield in January 2020, and was named XL.

[0433] II. Identification of the strain

[0434] Figure 1 The colony morphology diagram of Enterococcus faecium XL disclosed in the present application is shown in the figure. Figure 2 The cell morphology diagram of Enterococcus faecium XL disclosed in the present application is shown in the figure. Figure 1 and Figure 2 As shown in the figures, the morphological characteristics of the strain XL are as follows: the colony is grayish white, raised, fine and the periphery is neat, and the cell is oval arranged singly, in pairs or short chains.

[0435] The physiological and biochemical characteristics of the strain XL are as follows: anaerobic, growth temperature 20-60℃, optimum growth temperature 45℃, growth pH value range 2.0-8.0, and NaCl tolerance 0-8%.

[0436] According to the experimental method of “Bergey's Manual of Systematic Bacteriology”, the indicators such as gram staining, cell size and morphology, growth temperature, biochemical characteristics and the like are detected, and the phylogenetic tree is constructed according to the comparison results of the 16S rDNA gene sequence of the strain in GenBank (as shown in the figure). Figure 3 The strain XL is identified as a new strain and belongs to Enterococcus faecium.

[0437] III. Preservation of the strain

[0438] Preservation date: August 09, 2023

[0439] Preservation unit: China General Microbiological Culture Collection Center

[0440] Address of the preservation unit: No. 3, Yuanmingyuan West Road, Beijing City, China Academy of Microbiology

[0441] Preservation number: CGMCC No. 28133

[0442] Classification and naming: Enterococcus faecium.

[0443] IV. Physiological and biochemical properties of the strain

[0444] The partial physiological and biochemical characteristics of the Enterococcus faecium (the preservation number is CGMCC No. 28133) provided in the present application are shown in Table 1:

[0445] Table 1 Partial physiological and biochemical characteristics of Enterococcus faecium

[0446] Experimental items Results Experimental items Results Gram + Cellobiose + Oxidase - Raffinose - Contact enzyme - Xylose - V-P - Maltose + Glucose acid production + Lactose + Starch + Arabinose - Mannitol + Sucrose +

[0447] Note: "+" means positive reaction and "-" means negative reaction

[0448] According to the content of "Bergey's Mannual of Systematic Bacteriology", the strain XL belongs to Enterococcus according to its morphological characteristics and physiological and biochemical characteristics, and the phylogenetic tree is analyzed according to the alignment results of the 16S rDNA gene sequence of the bacteria in GenBank. The strain XL is a new bacteria, which is in a different branch from most Enterococcus bacteria, indicating that the bacteria has different growth characteristics, as shown in Table 1. Figure 3

[0449] Example 2: Performance evaluation of Enterococcus faecium XL provided by the application

[0450] (1) Growth and acid production ability evaluation

[0451] Enterococcus faecium XL was inoculated in the fermentation medium at an inoculation amount of 5%, and cultured at 40℃ for 2d, and the bacterial density, lactic acid content and pH value of the fermentation liquid were tested. The specific test data is shown in the following table:

[0452] Table 2 Cultivation of Enterococcus faecium

[0453] Culture time Bacterial density (cells / mL) Lactic acid content (mg / L) pH value 2d 4.8 x 10 8 ]]> 4020 3.8

[0454] (2) Evaluation of heavy component degradation ability

[0455] Enterococcus faecium XL was inoculated in the fermentation medium containing 2% crude oil at an inoculation amount of 5%, and cultured at 40℃ for 5d. According to the standard "SY / T5119-1995 Rock Soluble Organic Matter and Crude Oil Group Component Column Chromatography Analysis Method", the content of microbial asphaltene was reduced by 16%.

[0456] Table 3 Change of asphaltene before and after Enterococcus faecium XL

[0457] Culture time Before action (%) After action (%) 5d 28 12

[0458] (3) Evaluation of carbonate dissolution ability

[0459] ​Reference standard "QSH1020 2744-2019 Acidizing Plugging Removal Fluid Technical Requirements", evaluate the dissolution ability of microorganisms to calcium carbonate. Enterococcus faecalis XL was inoculated into the fermentation medium with marble blocks (20mm x 10mm x 10mm) at an inoculation amount of 5%, and the dissolution rate of marble was 23% after 6d of culture under the condition of 40℃.

[0460] Table 4 Dissolution rate of Enterococcus faecalis XL on marble

[0461]

[0462]

[0463] (4) Plugging removal performance evaluation

[0464] ① First, mix a certain proportion of quartz sand and crude oil uniformly according to a ratio of 30:1 to prepare oil sand, and use oil sand:oil sludge = 5:1 as the scale sample, wherein:

[0465] 20 The mass ratio of 20-40 mesh quartz sand to 40-80 mesh quartz sand is 20:80;

[0466] ② Fill the oil sand into the sand pack tube and age for 7d at 40℃, and perform a one-time water displacement to stabilize the pressure, and calculate the initial permeability K1 = 95 x 10 -3 μm 2 , the inlet end is A and the outlet end is B, and the displacement rate is 0.5mL / min;

[0467] ③ Stop the pump, take out 5cm of oil sand at the outlet end B after opening the sand pack tube, and then fill the scale sample;

[0468] ④ Perform a second water displacement to stabilize the pressure, and calculate the initial permeability K2 = 36 x 10 -3 μm 2 ;

[0469] ⑤ Inject 20mL of Enterococcus faecalis XL fermentation broth and fermentation nutrient mixture from the B end to the A end, and close for 5d at 40℃;

[0470] ⑥ Open the pump and perform three water displacements from the A end to the B end to stabilize the pressure, and calculate the permeability K3 = 109 x 10 -3 μm 2 after plugging removal, and the core permeability recovery rate is shown in Table 5:

[0471] Table 5 Enterococcus faecalis XL improves the core permeability

[0472]

[0473] (5) Physical model enhanced recovery evaluation

[0474] ①Core preparation: carbonate cementation core is filled, and air permeability is measured;

[0475] ②Vacuum extraction, saturation of simulated formation water, and measurement of core PV (pore volume);

[0476] ③Saturation of crude oil, water flooding for 7 days after core aging, water flooding to 92% of water content of produced water, and calculation of primary water flooding recovery rate;

[0477] ④Injection of 0.3 PV of Enterococcus faecalis XL fermentation broth and fermentation nutrient mixed solution, and culture for 15 days;

[0478] ⑤Secondary water flooding, water flooding to 100% of water content of produced liquid, and calculation of microbial enhanced oil recovery value of 18%.

[0479] As shown above, the performance of Enterococcus faecalis XL is as follows: bacterial concentration 4.8×10 8 6 / mL, lactic acid production 4020 mg / L, pH value 3.6; asphaltene content of crude oil is reduced by 16%; calcium carbonate corrosion rate is 23%; core permeability recovery rate is 115%; and physical simulation experiment enhanced oil recovery is greater than 18%.

[0480] Example 3: Application of the fermentation broth of Enterococcus faecalis XL provided by the application to microbial huff and puff of oil well A1 in a certain block of Shengli Oilfield.

[0481] Overview of oil well A1 in a certain block of Shengli Oilfield: formation temperature 60℃, formation water salinity 6870 mg / L, oil layer effective thickness 8 m, porosity 0.2, permeability 950×10 -3 μm 2 , underground crude oil viscosity 25 mPa·s, daily liquid 10 t / d, daily oil 0.7 t / d, and water content 93%.

[0482] The specific implementation steps are as follows:

[0483] According to the oil layer effective thickness, porosity, and treatment radius of the oil well, the total injection amount V of Enterococcus faecalis XL fermentation broth, fermentation medium, and water is calculated:

[0484] Total injection amount calculation formula: V=3.14R 2 Hфβ=75 m 3 , wherein:

[0485] V--total injection amount of Enterococcus faecalis XL fermentation broth, fermentation medium, and water, unit: m 3 ;

[0486] R--treatment radius, value: 5 m;

[0487] H--effective thickness, value: 8 m;

[0488] ф—porosity, its value is 0.2;

[0489] β—dosage coefficient, value is 0.6.

[0490] Take 7.5ml of fermentation broth of Enterococcus faecium XL 3 , fermentation medium 15m 3 , mix well and then inject 52.5m 3 water to obtain a mixed solution, wherein the volume concentration of the fermentation broth of Enterococcus faecium XL in the mixed solution is 10%;

[0491] The above 75m 3 The mixed liquid is injected into the formation through the annulus of the oil well casing, and then 50m of formation water is injected 3 , injection speed is 5m 3 / h, shut down the well for cultivation for 10 days and then reopen for production.

[0492] In this embodiment, the fermentation medium contains 10 g / L of carbon source, 5 g / L of nitrogen source, 0.5 g / L of phosphorus source, 1 g / L of diammonium hydrogen citrate, 0.1 g / L of magnesium sulfate, and 1 g / L of sodium acetate.

[0493] The carbon source of the fermentation medium is sucrose, the nitrogen source is corn steep liquor, and the phosphorus source is dipotassium hydrogen phosphate. The fermentation conditions of Enterococcus faecium XL are as follows: inoculation amount 1%, inoculation age 10 hours, temperature 35°C, and static culture for 24 hours.

[0494] In another embodiment, the fermentation medium comprises: 15 g / L carbon source, 8 g / L nitrogen source, 1 g / L phosphorus source, 1.5 g / L diammonium hydrogen citrate, 0.3 g / L magnesium sulfate, and 3 g / L sodium acetate, wherein:

[0495] The carbon source of the fermentation medium is sucrose, the nitrogen source is corn steep liquor and yeast powder in equal mass ratio, and the phosphorus source is dipotassium hydrogen phosphate.

[0496] The fermentation conditions of Enterococcus faecium XL are as follows: inoculation amount 3%, inoculation age 12 hours, temperature 40° C., and static culture for 30 hours.

[0497] In another embodiment, the fermentation medium comprises: 20 g / L carbon source, 10 g / L nitrogen source, 2 g / L phosphorus source, 2 g / L diammonium hydrogen citrate, 0.5 g / L magnesium sulfate, and 5 g / L sodium acetate.

[0498] The carbon source of the fermentation medium is sucrose, glucose, dextrin and molasses in equal mass ratio; the nitrogen source is corn steep liquor, soybean cake powder, peptone, beef extract and yeast powder in equal mass ratio; and the phosphorus source is dipotassium hydrogen phosphate, diammonium hydrogen phosphate and disodium hydrogen phosphate in equal mass ratio.

[0499] The fermentation conditions of Enterococcus faecium XL are as follows: inoculation amount 5%, inoculation age 15h, temperature 45℃, and static culture for 48h.

[0500] Test results: the liquid and oil production of the well are increased, the peak daily liquid is increased from 10m3 to 22m3, the daily oil is increased from 0.7t to 4.0t, the water content is decreased from 93% to 82%, the cumulative oil increment is 750t, the effective period reaches 280d, the organic acid produced by the growth and metabolism of Enterococcus faecium XL plays a function of plugging removal on the oil well, the single-well productivity is finally improved, and the field test effect is good.

[0501] Example 4: The application of Enterococcus faecium XL microbial agent provided by the application in microbial single-well huff and puff of an oil well B3 in a certain block of Shengli Oilfield.

[0502] The oil well B3 in a certain block of Shengli Oilfield is as follows: formation temperature 52℃, formation water salinity 670mg / L, oil layer effective thickness 16m, porosity 0.12, permeability 360x10 -3 μm 2 , underground crude oil viscosity 630mPa·s, daily liquid 12t / d, daily oil 0.6t / d, and water content 95%.

[0503] The specific implementation steps are as follows:

[0504] According to the oil layer effective thickness, porosity and treatment radius of the oil well, the total injection amount V of Enterococcus faecium XL fermentation liquid, fermentation medium and water is calculated:

[0505] The total injection amount calculation formula is: V=3.14R 2 Hфβ=392m 3 , wherein:

[0506] V is the total injection amount of Enterococcus faecium XL fermentation liquid, fermentation medium and water, and the unit is m 3 ;

[0507] R is the treatment radius, and the value is 8.5m;

[0508] H is the effective thickness, and the value is 16m;

[0509] ф is the porosity, and the value is 0.12;

[0510] β is the dosage coefficient, and the value is 0.9.

[0511] The first slug injection: 58.8m 3 of Enterococcus faecium XL fermentation liquid, 88.2m 3 of Enterococcus faecium XL fermentation medium, and 245m 3 of water are mixed to obtain a mixed liquid, and the volume concentration of Enterococcus faecium XL fermentation liquid in the mixed liquid is 15%;

[0512] The 392m 3 The mixed solution is injected into the stratum from the oil well casing annulus, and then stratum water is injected 60m 3 at a speed of 8m 3 / h, and the well is closed for 10d.

[0513] The fermentation medium comprises 10g / L of carbon source, 5g / L of nitrogen source, 0.5g / L of phosphorus source, 1.0g / L of diammonium hydrogen citrate, 0.1g / L of magnesium sulfate, and 1.0g / L of sodium acetate.

[0514] The carbon source of the fermentation medium is glucose, the nitrogen source is soybean meal, and the phosphorus source is diammonium hydrogen phosphate.

[0515] The fermentation conditions of the Enterococcus faecium XL are as follows: inoculation amount 1%, inoculation age 10h, temperature 35℃, and static culture for 24h.

[0516] In another embodiment, the fermentation medium comprises 15g / L of carbon source, 8g / L of nitrogen source, 1.2g / L of phosphorus source, 1.5g / L of diammonium hydrogen citrate, 0.3g / L of magnesium sulfate, and 2.0g / L of sodium acetate.

[0517] The carbon source of the fermentation medium is glucose, the nitrogen source is soybean meal, and the phosphorus source is diammonium hydrogen phosphate.

[0518] The fermentation conditions of the Enterococcus faecium XL are as follows: inoculation amount 3%, inoculation age 12h, temperature 40℃, and static culture for 30h.

[0519] In yet another embodiment, the fermentation medium comprises 20g / L of carbon source, 10g / L of nitrogen source, 2g / L of phosphorus source, 2g / L of diammonium hydrogen citrate, 0.5g / L of magnesium sulfate, and 5.0g / L of sodium acetate.

[0520] The carbon source of the fermentation medium is glucose, the nitrogen source is soybean meal, and the phosphorus source is diammonium hydrogen phosphate.

[0521] The fermentation conditions of the Enterococcus faecium XL are as follows: inoculation amount 5%, inoculation age 15h, temperature 45℃, and static culture for 48h.

[0522] The second slug injection: first, water is injected 20m 3 , and then

[0523] 50m 3 of the red sphericae fermentation liquid, 50m 3 of the activator are taken respectively, and after being uniformly mixed, an appropriate amount of water is injected thereinto to obtain a mixed solution, wherein the volume concentration of the red sphericae fermentation liquid in the mixed solution is 5%, the mixed solution is injected into the stratum from the oil well casing annulus, and then stratum water is injected 80m 3 at a speed of 10m 3 / h, the well is shut down for 10 days and then opened for production.

[0524] The activator is glucose 10 g / L, proteose peptone 3 g / L, sodium nitrate 5 g / L, disodium hydrogen phosphate 1 g / L, potassium dihydrogen phosphate 0.5 g / L, small molecule peptide 0.3 g / L, and the balance is water.

[0525] In another embodiment, the activator is glucose 12 g / L, proteose peptone 4 g / L, sodium nitrate 8 g / L, disodium hydrogen phosphate 1.2 g / L, potassium dihydrogen phosphate 0.8 g / L, small molecule peptide 0.4 g / L, and the balance is water.

[0526] In yet another embodiment, the activator is glucose 15 g / L, proteose peptone 5 g / L, sodium nitrate 10 g / L, disodium hydrogen phosphate 1.5 g / L, potassium dihydrogen phosphate 1 g / L, small molecule peptide 0.5 g / L, and the balance is water.

[0527] Test results: the liquid and oil production of the well both increased, the peak daily liquid increased from 12 t to 30 t, the daily oil increased from 0.6 t to 6 t, the water content decreased from 95% to 80%, the underground crude oil viscosity decreased from 630 mPa·s to 120 mPa·s, the crude oil viscosity reduction rate reached 81%, the cumulative oil increase was 1650 t, the effective period reached 500 d, the organic acids produced by the growth and metabolism of Enterococcus faecalis XL played a role in plugging removal of the oil well, and the biosurfactant produced by the growth and metabolism of the biological emulsifying bacteria realized the emulsification and oil washing effect of the crude oil, thereby improving the single well productivity.

[0528] Example 5: The application provides an application of the Enterococcus faecalis XL fermentation liquor in microbial oil displacement in a D 2 block of Shengli Oilfield.

[0529] The well group profile of the D 2 block of Shengli Oilfield is as follows: formation temperature 48℃, formation water salinity 12500 mg / L, oil layer effective thickness 5.4 m, porosity 0.30, permeability 930 x 10 -3 μm 2 , underground crude oil viscosity 60 mPa·s, 1 injection and 3 production, block daily liquid production 34 m3, daily oil production 2.9 t, and comprehensive water content 92.2%.

[0530] The specific implementation steps are as follows:

[0531] An appropriate amount of Enterococcus faecalis fermentation liquor, fermentation medium and water are weighed and mixed uniformly to obtain a mixed liquor, then the mixed liquor is injected into a water injection well, then normal water injection is performed for 5 days, the Enterococcus faecalis is pushed into the deep part of the oil reservoir, and the injection is stopped for 8 days for cultivation, so that the Enterococcus faecalis is fully reproduced and metabolized in the oil reservoir, then normal water injection is performed for 10 days, and one cycle of injection is completed.

[0532] The total injection amount of Enterococcus faecium fermentation liquor, fermentation medium and water is 500m 3 ;

[0533] V 屎肠球菌发酵液 :V 发酵培养基 =1:5;

[0534] The volume concentration of Enterococcus faecium fermentation liquor in the mixed liquor is 5%.

[0535] The fermentation medium refers to 10g / L of carbon source, 5g / L of nitrogen source, 0.5g / L of phosphorus source, 11.0g / L of diammonium hydrogen citrate, 0.1g / L of magnesium sulfate, 1g / L of sodium acetate, and the rest is water.

[0536] The carbon source is dextrin, the nitrogen source is peptone, and the phosphorus source is disodium hydrogen phosphate.

[0537] The fermentation conditions of Enterococcus faecium XL are as follows: inoculation amount 1%, inoculation age 10h, temperature 35℃, and static culture 24h.

[0538] In another embodiment, the application comprises the following steps:

[0539] An appropriate amount of Enterococcus faecium fermentation liquor, fermentation medium and water are weighed respectively, mixed uniformly to obtain a mixed liquor, and then the mixed liquor is injected into a water injection well, then normal water injection is carried out for 10d, Enterococcus faecium is pushed into the deep part of the oil reservoir, and the injection is stopped for culture for 20d, so that Enterococcus faecium is fully propagated and metabolized in the oil reservoir, and then normal water injection is carried out for 15d, to complete one cycle of injection, wherein:

[0540] The total injection amount of Enterococcus faecium fermentation liquor, fermentation medium and water is 1000m 3 ;

[0541] V 屎肠球菌发酵液 :V 发酵培养基 =1:8;

[0542] The volume concentration of Enterococcus faecium fermentation liquor in the mixed liquor is 10%.

[0543] The fermentation medium refers to 15g / L of carbon source, 8g / L of nitrogen source, 1.2g / L of phosphorus source, 1.5g / L of diammonium citrate, 0.3g / L of magnesium sulfate, 2.0g / L of sodium acetate, and the rest is water.

[0544] The carbon source is dextrin, the nitrogen source is peptone, and the phosphorus source is disodium hydrogen phosphate.

[0545] The fermentation conditions of Enterococcus faecium XL are as follows: inoculation amount 3%, inoculation age 12h, temperature 40℃, and static culture 30h.

[0546] In yet another embodiment, the application comprises the following steps:

[0547] Weigh appropriate amounts of Enterococcus faecium fermentation liquid, fermentation medium and water respectively, mix them evenly to obtain a mixed solution, and then inject the mixed solution into the water injection well. Then, inject water normally for 8 days to push Enterococcus faecium into the deep part of the oil reservoir. Stop injection and culture for 12 days to allow Enterococcus faecium to fully reproduce and metabolize in the oil reservoir. Then, inject water normally for 12 days to complete one injection cycle.

[0548] The total injection volume of Enterococcus faecium fermentation liquid, fermentation medium and water is 800m 3 ;

[0549] V 屎肠球菌发酵液 :V 发酵培养基 =1:6;

[0550] The volume concentration of the Enterococcus faecium fermentation liquid in the mixed liquid is 8%.

[0551] The fermentation medium comprises 20 g / L of carbon source, 10 g / L of nitrogen source, 2 g / L of phosphorus source, 2 g / L of diammonium hydrogen citrate, 0.5 g / L of magnesium sulfate, 5.0 g / L of sodium acetate, and the balance is water.

[0552] The carbon source is dextrin, the nitrogen source is peptone, and the phosphorus source is disodium hydrogen phosphate.

[0553] The fermentation conditions of Enterococcus faecium XL are as follows: inoculation amount 5%, inoculation age 15 hours, temperature 45° C., and static culture for 48 hours.

[0554] Test Results: After the test, both liquid and oil production increased in the well group, as shown in Table 6. The well group's liquid volume increased by 21 tons / day, production increased by 5.7 tons / day, and water cut decreased by 7.3 percentage points, resulting in a cumulative increase in oil production of 1,800 tons. The effective period lasted 560 days. The field test demonstrated positive results by utilizing the organic acids produced by the growth and metabolism of Enterococcus faecium XL to enhance formation permeability, ultimately increasing production in the reservoir well group.

[0555] Table 6 Effects of well groups after on-site implementation

[0556]

[0557] Example 6: Application of the Enterococcus faecium XL fermentation broth provided by the present invention in microbial oil recovery in the F3 block well group of Shengli Oilfield.

[0558] Overview of the well group in the F3 block of Shengli Oilfield: formation temperature 50°C, formation water salinity 52500 mg / L, effective oil layer thickness 11 m, porosity 0.26, permeability 780×10 -3 μm 2 The underground crude oil viscosity is 506 mPa·s, the well group is 1 injection and 6 production, the block produces 80 tons of liquid and 7.8 tons of oil per day, and the comprehensive water content is 90.8%.

[0559] The specific implementation steps are as follows:

[0560] The specific steps of the application include:

[0561] The first slug: an appropriate amount of Enterococcus faecium fermentation liquor, fermentation medium and water are weighed respectively, mixed uniformly to obtain a mixed liquor, and then the mixed liquor is injected into the injection well, wherein:

[0562] The total injection amount of the Enterococcus faecium fermentation liquor, the fermentation medium and the water is 800m 3 ;

[0563] V 屎肠球菌发酵液 :V 发酵培养基 =1:7;

[0564] The volume concentration of the Enterococcus faecium fermentation liquor in the mixed liquor is 8%;

[0565] The fermentation medium refers to 10g / L of carbon source, 5g / L of nitrogen source, 0.5g / L of phosphorus source, 11.0g / L of diammonium hydrogen citrate, 0.1g / L of magnesium sulfate, 1g / L of sodium acetate, and the pH value is 6.5, and the rest is water.

[0566] The carbon source is molasses, the nitrogen source is beef extract, and the phosphorus source is diammonium hydrogen phosphate.

[0567] The fermentation conditions of the Enterococcus faecium XL are as follows: inoculation amount 1%, inoculation age 10h, temperature 35℃, and static culture for 24h.

[0568] The second slug is injected: first, water is injected for 4d, then an appropriate amount of bio-emulsifier fermentation liquor, activator and water are weighed respectively, mixed uniformly to obtain a mixed liquor, the mixed liquor is injected into the injection well, then normal water injection is performed for 12d, the injection is stopped for culture for 15d, then normal water injection is performed for 18d, and one cycle of injection is completed, wherein:

[0569] V 生物乳化剂发酵液 :V 激活剂 =1:3;

[0570] The total injection amount of the bio-emulsifier fermentation liquor, the activator and the water is 600m 3 ;

[0571] The volume concentration of the bio-emulsifier fermentation liquor in the mixed liquor is 12%;

[0572] The bio-emulsifier fermentation liquor is Rhodococcus fermentation liquor;

[0573] The activator refers to glucose 10-15 g / L, proteose peptone 3-5 g / L, sodium nitrate 5-10 g / L, disodium hydrogen phosphate 1-1.5 g / L, potassium dihydrogen phosphate 0.5-1 g / L, growth factor 0.3-0.5 g / L, and pH value is 6.0-8.0;

[0574] The growth factor is one of vitamin, small molecule peptide, amino acid, aromatic hydrocarbon, alkane and xylitol.

[0575] In another embodiment, the specific steps of the application include:

[0576] The first plug: an appropriate amount of Enterococcus faecium fermentation liquor, fermentation medium and water are weighed respectively, mixed uniformly to obtain a mixed liquor, and then the mixed liquor is injected into the injection well, wherein:

[0577] The total injection amount of Enterococcus faecium fermentation liquor, fermentation medium and water is 1000 m 3 ;

[0578] V 屎肠球菌发酵液 :V 发酵培养基 =1:8;

[0579] The volume concentration of Enterococcus faecium fermentation liquor in the mixed liquor is 10%;

[0580] The second plug is injected: first, water is injected for 5 d, then an appropriate amount of bio-emulsifier fermentation liquor, activator and water are weighed respectively, mixed uniformly to obtain a mixed liquor, the mixed liquor is injected into the injection well, then normal water injection is carried out for 15 d, injection is stopped for 25 d, and then normal water injection is carried out for 20 d, to complete a cycle of injection, wherein:

[0581] V 生物乳化剂发酵液 :V 激活剂 =1:5;

[0582] The total injection amount of bio-emulsifier fermentation liquor, activator and water is 800 m 3 ;

[0583] The volume concentration of bio-emulsifier fermentation liquor in the mixed liquor is 15%;

[0584] The bio-emulsifier fermentation liquor is Bacillus tekila fermentation liquor;

[0585] The activator refers to glucose 12 g / L, proteose peptone 3.5 g / L, sodium nitrate 7.5 g / L, disodium hydrogen phosphate 1.25 g / L, potassium dihydrogen phosphate 0.75 g / L, growth factor 0.4 g / L, and pH value is 7.0;

[0586] The growth factor is vitamin.

[0587] In yet another embodiment, the specific steps of the application include:

[0588] The first slug: the appropriate amount of Enterococcus faecium fermentation liquor, fermentation medium and water are weighed respectively, mixed uniformly to obtain a mixed liquor, and then the mixed liquor is injected into the injection well, wherein:

[0589] The total injection amount of the Enterococcus faecium fermentation liquor, the fermentation medium and water is 500m 3 ;

[0590] V 屎肠球菌发酵液 :V 发酵培养基 =1:5;

[0591] The volume concentration of the Enterococcus faecium fermentation liquor in the mixed liquor is 5%;

[0592] The second slug is injected: first, water is injected for 3d, then the appropriate amount of bio-emulsifier fermentation liquor, activator and water are weighed respectively, mixed uniformly to obtain a mixed liquor, the mixed liquor is injected into the injection well, then normal water injection is carried out for 8d, injection is stopped for 10d for culture, and then normal water injection is carried out for 15d, to complete a cycle of injection, wherein:

[0593] V 生物乳化剂发酵液 :V 激活剂 =1:2;

[0594] The total injection amount of the bio-emulsifier fermentation liquor, the activator and water is 300m 3 ;

[0595] The volume concentration of the bio-emulsifier fermentation liquor in the mixed liquor is 10%;

[0596] The bio-emulsifier fermentation liquor is Arthrobacter fermentation liquor;

[0597] The activator refers to 10g / L of glucose, 5g / L of proteose peptone, 5g / L of sodium nitrate, 1g / L of disodium hydrogen phosphate, 0.5g / L of potassium dihydrogen phosphate, 0.3g / L of growth factor, and the pH value is 6.0;

[0598] The growth factor is a small molecule peptide.

[0599] Test results: the liquid production and oil production of the well group are both increased after the test, and the specific conditions are shown in Table 7, wherein the liquid amount of the well group is increased by 42t / d, the production is increased by 13.3t / d, the water content is reduced by 7.3 percentage points, the cumulative oil increase is 3260t, and the effective period reaches 380d. The organic acid produced by the growth and metabolism of Enterococcus faecium XL dissolves the formation, improves the formation permeability, and realizes the emulsification and oil washing effect of crude oil through the bio-surfactant produced by the growth and metabolism of the bio-emulsifying bacteria to synergistically improve the production of the well group. The field test effect is good.

[0600] Table 7 Effect of well group after field implementation

[0601]

[0602] Example 7

[0603] The same as Example 6, the only difference is that:

[0604] 1. The fermentation medium refers to carbon source 15 g / L, nitrogen source 8 g / L, phosphorus source 1.2 g / L, diammonium hydrogen citrate 1.5 g / L, magnesium sulfate 0.3 g / L, sodium acetate 2.0 g / L, pH 6.8, and the balance is water.

[0605] The carbon source is molasses, the nitrogen source is beef extract, and the phosphorus source is diammonium hydrogen phosphate.

[0606] The fermentation conditions of Enterococcus faecium XL are inoculation amount 3%, inoculation age 12 h, temperature 40℃, and static culture for 30 h.

[0607] 2. The bio-emulsified fermentation broth is Bacillus pallidus fermentation broth.

[0608] The activator refers to glucose 12 g / L, proteose peptone 3.5 g / L, sodium nitrate 7 g / L, disodium hydrogen phosphate 1.2 g / L, potassium dihydrogen phosphate 0.7 g / L, small molecule peptide 0.35 g / L, pH 6.0, and the balance is water.

[0609] Example 8

[0610] The same as Example 6, the only difference is that:

[0611] 1. The fermentation medium refers to carbon source 20 g / L, nitrogen source 10 g / L, phosphorus source 2 g / L, diammonium hydrogen citrate 2 g / L, magnesium sulfate 0.5 g / L, sodium acetate 5.0 g / L, pH 7.5, and the balance is water.

[0612] The carbon source is molasses, the nitrogen source is beef extract, and the phosphorus source is diammonium hydrogen phosphate.

[0613] The fermentation conditions of Enterococcus faecium XL are inoculation amount 5%, inoculation age 15 h, temperature 45℃, and static culture for 48 h.

[0614] 2. The bio-emulsified fermentation broth is Bacillus tequilensis fermentation broth.

[0615] The activator is glucose 10 g / L, proteose peptone 3 g / L, sodium nitrate 5 g / L, disodium hydrogen phosphate 1 g / L, potassium dihydrogen phosphate 0.5 g / L, amino acid 0.3 g / L, pH 7.2, and the balance is water.

[0616] Example 9

[0617] The same as Example 6, the only difference is that:

[0618] 1. The fermentation medium comprises 16 g / L of carbon source, 6 g / L of nitrogen source, 0.8 g / L of phosphorus source, 1.6 g / L of dihydrogen ammonium citrate, 0.5 g / L of magnesium sulfate, 3.0 g / L of sodium acetate, and the rest is water, and the pH value is 7.0.

[0619] The carbon source is dextrin, the nitrogen source is soybean meal, and the phosphorus source is dihydrogen ammonium phosphate.

[0620] The fermentation conditions of Enterococcus faecium XL are as follows: inoculation amount 4%, inoculation age 13 h, temperature 36℃, and static culture 38 h.

[0621] 2. The biological emulsified fermentation liquid is a red ball fermentation liquid.

[0622] The activator comprises 12 g / L of glucose, 4 g / L of proteose peptone, 8 g / L of sodium nitrate, 1.2 g / L of di-sodium hydrogen phosphate, 0.8 g / L of potassium dihydrogen phosphate, 0.4 g / L of amino acid, and the rest is water, and the pH value is 7.5.

[0623] The above describes the embodiments of the present application in detail. However, the present application is not limited to the above embodiments, and various changes can be made within the knowledge of those skilled in the art without departing from the spirit of the present application.

Claims

1. A strain of Enterococcus faecium XL, characterized in that, The preservation number of which is CGMCC No. 28133.

2. An inoculant characterized in that, The effective component of which is Enterococcus faecium as claimed in claim 1.

3. A fermentation broth, characterized in that, The liquid which is metabolized by Enterococcus faecium as claimed in claim 1 in a fermentation medium, wherein: The active component of the fermentation liquid is Enterococcus faecium as claimed in claim 1.

4. A fermentation broth as claimed in claim 3, wherein, The fermentation medium comprises: carbon source at least 10 g / L, nitrogen source at least 2 g / L, phosphorus source at least 0.2 g / L, inorganic salt at least 0.2 g / L, pH value 6.5-7.5, the rest is water, and / or The carbon source is at least one of sucrose, glucose, dextrin, molasses, galactose, arabinose, melibiose, xylose, maltose, and / or The nitrogen source is at least one of corn steep liquor, soybean meal, peptone, beef extract, yeast powder, urea, ammonium chloride, and / or The phosphorus source is at least one of dipotassium hydrogen phosphate, diammonium hydrogen phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, ammonium dihydrogen phosphate, and / or The inorganic salt is at least one of magnesium sulfate, sodium acetate, potassium sulfate, potassium chloride, sodium chloride, magnesium chloride.

5. A fermentation broth as claimed in claim 3, wherein, The fermentation conditions of Enterococcus faecium are as follows: inoculation amount is at least 0.2% V / V inoculation age is at least 5 hours, fermentation temperature is 35-45℃, and static culture is at least 12 hours. Preferably, the Enterococcus faecalis fermentation conditions are: inoculum 1-5% V / V , inoculum age 10-15 h, fermentation temperature 35-45 °C, static culture 24-48 h.

6. Application of substance I and / or substance II and / or substance III in oil exploitation, wherein: Substance I is Enterococcus faecium as claimed in claim 1; Substance II is the bacterial agent as claimed in claim 2; Substance III is the fermentation liquid as claimed in claim 3.

7. Application of substance I and / or substance II and / or substance III in microbial oil production of oil well in middle-low permeability water flooding reservoir, wherein: Substance I is Enterococcus faecium as claimed in claim 1; Substance II is the bacterial agent as claimed in claim 2; Substance III is the fermentation liquid as claimed in claim 3.

8. Application of substance I and / or substance II and / or substance III in microbial single well huff and puff of oil well in middle-low permeability water flooding reservoir, wherein: Substance I is Enterococcus faecium as claimed in claim 1; Substance II is the bacterial agent as claimed in claim 2; Substance III is the fermentation liquid as claimed in claim 3.

9. Application of substance I and / or substance II and / or substance III in microbial oil displacement in middle-low permeability water flooding reservoir, wherein: Substance I is Enterococcus faecium as claimed in claim 1; Substance II is the bacterial agent as claimed in claim 2; Substance III is the fermentation liquid as claimed in claim 3.

10. A product characterized by, The active component of the product is substance I and / or substance II and / or substance III; Substance I is Enterococcus faecium as claimed in claim 1; Substance II is the bacterial agent as claimed in claim 2; Substance III is the fermentation liquid as claimed in claim 3; The function of the product is (a) or (b) or (c) or (d): (a) oil exploitation; (b) microbial oil production; (c) microbial single well huff and puff; (d) microbial oil displacement.

11. A method of oil recovery, characterized by, Comprise the following steps: Substance I and / or substance II and / or substance III are added in the process of oil production; Substance I is Enterococcus faecium as claimed in claim 1; Substance II is the bacterial agent as claimed in claim 2; Substance III is the fermentation liquid as claimed in claim 3.

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