Biological paraffin inhibitor composite strain culture system and special formula for oil field

Abstract

The invention discloses a biological paraffin inhibitor composite strain culture system and a special formula for an oil field, and belongs to the field of petroleum microorganisms. The composite strain applied in the invention has good passage stability, and after 12 generations of passage, the paraffin reduction rate of the solid paraffin can be stabilized at 19.6%; and the wax reduction rate of fermentation at a shake flask level reaches 35-45%. The method for fermenting and degrading the paraffin is simple and easy to operate, low in culture medium cost and suitable for production and application in the petroleum industry.

Description

Technical Field

[0001] This invention relates to a biological wax-resistant compound microbial culture system and an oilfield-specific formula, belonging to the field of petroleum microbial technology. Background Technology

[0002] Pipeline flow safety is a critical issue in crude oil transportation. In buried or subsea oil gathering pipelines located in frigid regions, the low temperatures of the surrounding soil or seawater cause significant heat loss during transport, leading to a gradual decrease in crude oil temperature. When the crude oil temperature falls below its wax precipitation point, the wax component begins to precipitate, slowly accumulating and growing, eventually depositing on the pipeline's inner wall. As the wax deposit increases, it reduces the pipeline's effective radius, increases flow resistance, and in severe cases, can cause blockages, resulting in substantial economic losses. Globally, addressing problems caused by wax deposition during crude oil extraction, transportation, storage, and processing costs hundreds of millions of dollars annually.

[0003] Currently, the main methods for controlling wax deposition in oil wells include physical methods, chemical methods, and biological methods.

[0004] Physical methods, taking thermal dewaxing as an example, involve increasing the temperature of the oil flow and oil pipes through thermodynamic means to remove wax deposits. These methods include steam dewaxing, thermochemical dewaxing, and hot fluid circulation dewaxing. However, due to their relatively high cost and low efficiency, they can only be used as supplementary dewaxing methods.

[0005] Chemical methods involve adding chemicals to the pipeline medium to prevent wax deposition and gelation. Based on their mechanisms of action, these can be categorized into three types: chemical wax inhibitors, pour point depressants, and paraffin dispersants. Chemicals that affect the wax precipitation temperature of crude oil are generally called wax inhibitors. Chemicals that affect the pour point of crude oil are called pour point depressants. Since both types of chemicals work by influencing the wax crystallization process, there is significant overlap between them. Paraffin dispersants are surfactants that work by adsorbing onto the pipeline wall and wax crystals. However, this method has drawbacks such as high dosage, high cost, and poor safety and environmental performance.

[0006] Conventional microbial methods utilize microbial wax-removing agents to prevent wax formation in oil wells. Microorganisms, after adhering to and growing on the surfaces of metals, clays, or minerals, form a dense protective microbial film. This film adheres to the pipe walls of oil well equipment, increasing the wettability of the pipe surface and significantly improving crude oil flow, thus preventing wax crystallization. Furthermore, through microbial metabolism, secondary metabolites from alkanes are produced, such as lipopeptides, glycolipids, fatty acids, and lipid bodies—surfactants with biological properties. Their emulsifying effect increases lubrication during oil extraction. Microorganisms can also convert n-alkanes in crude oil into isoalkanes, distorting wax crystal formation and preventing crystallization. Simultaneously, the small molecules produced by microorganisms, such as organic acids, ethanol, and acetaldehyde, greatly increase the solubility of crude oil. The generated gases, such as carbon dioxide and methane, also reduce the viscosity of crude oil, effectively shielding crystal nuclei, preventing further crystal growth, and effectively preventing paraffin deposition.

[0007] Given the enormous potential of microbial methods in the application of wax inhibitors, the development of more efficient, environmentally friendly, and low-cost microbial wax inhibitors is urgently needed. This will not only help meet market demand for wax inhibitors but also promote the sustainable development of related petroleum industries. Summary of the Invention

[0008] To address the aforementioned problems, this invention combines Alcanivorax dieselolei (accession number CGMCC No. 1.8628) and Alcanivorax jadensis (accession number CGMCC No. 1.16108) to obtain a compound bacterial agent with high efficiency in degrading paraffin. Based on this, the contents of glucose, sodium nitrate, ammonium sulfate, potassium dihydrogen phosphate, magnesium sulfate, ferrous sulfate, and calcium chloride in the fermentation medium were optimized.

[0009] This invention provides a microbial inoculant.

[0010] In one embodiment of the present invention, the microbial agent comprises *Alcanivorax dieselolei* (CGMCC No. 1.8628) and *Alcanivorax jadensis* (CGMCC No. 1.16108). The ratio of *Alcanivorax dieselolei* CGMCC No. 1.8628 to *Alcanivorax jadensis* CGMCC No. 1.16108 is 1–2:1–2.

[0011] In one embodiment of the present invention, the viable count in the microbial agent reaches 1 to 3 × 10⁻⁶. 6 CFU / g or CFU / mL.

[0012] In one embodiment of the present invention, the fermentation system contains 1-20 g / L glucose, 1.000-3.000 g / L sodium nitrate, 1.000-3.000 g / L ammonium sulfate, 0.000-4.000 g / L dipotassium hydrogen phosphate, 0-0.025 g / L magnesium sulfate, 0-0.080 g / L potassium chloride, 0-0.025 g / L ferrous sulfate, and 0-0.008 g / L calcium chloride.

[0013] In one embodiment of the present invention, the fermentation system contains 1-20 g / L glucose, 1.000-3.000 g / L sodium nitrate, 1.000-3.000 g / L ammonium sulfate, 1.000-4.000 g / L dipotassium hydrogen phosphate, 0-0.025 g / L magnesium sulfate, 0.010-0.080 g / L potassium chloride, 0.010-0.025 g / L ferrous sulfate, and 0.002-0.008 g / L calcium chloride.

[0014] In one embodiment of the present invention, the concentration of glucose is preferably 10 g / L, the concentration of sodium nitrate is preferably 2.000 g / L, the concentration of ammonium sulfate is preferably 2.000 g / L, the concentration of dipotassium hydrogen phosphate is preferably 2.000 g / L, the concentration of magnesium sulfate is preferably 0.700 g / L, the concentration of potassium chloride is preferably 0.050 g / L, the concentration of ferrous sulfate is preferably 0.015 g / L, and the concentration of calcium chloride is preferably 0.004 g / L.

[0015] This invention provides a method for degrading paraffin, wherein the microbial agent described above is inoculated into a solid paraffin culture medium for fermentation; wherein the concentration of solid paraffin is 10-50 g / L.

[0016] In one embodiment of the present invention, the solid paraffin culture medium further contains: 1-20 g / L glucose concentration, 1-3 g / L sodium nitrate concentration, 1-3 g / L ammonium sulfate concentration, 1-4 g / L dipotassium hydrogen phosphate concentration, 0-0.9 g / L magnesium sulfate concentration, 0-0.08 g / L potassium chloride concentration, 0-0.025 g / L ferrous sulfate concentration, and 0-0.008 g / L calcium chloride concentration, with a pH of 6-10.

[0017] In one embodiment of the present invention, the solid paraffin culture medium contains glucose at a concentration of 10 g / L, sodium nitrate at a concentration of 2 g / L, ammonium sulfate at a concentration of 2 g / L, dipotassium hydrogen phosphate at a concentration of 2 g / L, magnesium sulfate at a concentration of 0.9 g / L, potassium chloride at a concentration of 0.05 g / L, ferrous sulfate at a concentration of 0.015 g / L, calcium chloride at a concentration of 0.004 g / L, and pH 8.

[0018] In one embodiment of the present invention, the fermentation time is 168-192 h and the fermentation temperature is 25-35 °C; preferably, the fermentation time is 168 h, the fermentation temperature is 30 °C, and the solid paraffin concentration is 20 g / L.

[0019] In one embodiment of the present invention, the concentration of the bacterial agent is 1 to 3 × 10⁻⁶. 6 The CFU / mL is preferably 20 g / L.

[0020] This invention also protects the application of the aforementioned anti-wax bacteria culture system in the production of biological anti-wax agents.

[0021] Beneficial effects

[0022] (1) The culture system of the present invention enables the composite strain to have good passage stability. After 12 passages, its solid paraffin degradation ability can be stabilized at 19.6%; the degradation rate of solid paraffin at the shake flask level reaches 43.0%.

[0023] (2) The method for fermenting and degrading solid paraffin in this invention is simple and easy to operate, the culture medium is inexpensive, and the obtained microbial cells are suitable for paraffin prevention applications in oil fields. Attached Figure Description

[0024] Figure 1 This is a graph showing the relationship between the initial pH of fermentation and the degradation of solid paraffin.

[0025] Figure 2 A graph showing the relationship between the initial concentration of solid paraffin during fermentation and the degradation of solid paraffin.

[0026] Figure 3 The graph shows the change in wax reduction rate during the horizontal fermentation process of the compound bacteria in a shake flask. Detailed Implementation

[0027] The present invention will be further described below with reference to the accompanying drawings and specific embodiments. These embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, where specific conditions are not specified, are generally performed under conventional conditions in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those familiar with the art.

[0028] The culture media involved in the following examples are as follows:

[0029] Slant culture medium: ammonium sulfate 8.00 g / L, ammonium nitrate 1.00 g / L, potassium dihydrogen phosphate 1.00 g / L, dipotassium hydrogen phosphate 1.00 g / L, magnesium sulfate 0.20 g / L, ferric chloride 0.03 g / L, calcium chloride 0.02 g / L, sodium chloride 5.00 g / L, glucose 20.00 g / L, agar 20.00 g / L. Sterilize at 115℃ for 30 min.

[0030] Seed culture medium (g / L): sodium chloride 19.59 g / L, magnesium chloride 5.90 g / L, magnesium sulfate 3.24 g / L, calcium chloride 1.80 g / L, potassium chloride 0.55 g / L, potassium nitrate 2.00 g / L, dipotassium hydrogen phosphate 0.126 g / L, ammonium nitrate 2.00 g / L, sodium acetate 2.00 g / L, yeast extract 0.50 g / L, peptone 0.50 g / L, glucose 0.20 g / L, sodium citrate 1.00 g / L, ferrous sulfate 0.01 g / L; sterilized at 115℃ for 30 min.

[0031] Fermentation medium (g / L): glucose 1g / L, sodium nitrate 1.500g / L, ammonium sulfate 1.500g / L, dipotassium hydrogen phosphate 1.000g / L, magnesium sulfate 0.500g / L, potassium chloride 0.050g / L, ferrous sulfate 0.010g / L, calcium chloride 0.002g / L, solid paraffin 10g / L.

[0032] Strains:

[0033] Alcanivorax dieselolei (CGMCC NO.1.8628), Alcanivorax di eselolei (CGMCC NO.1.3690), and Alcanivorax jadensis (CGMCC NO.1.16108) were purchased from the China General Microbiological Culture Collection Center.

[0034] The calculation methods involved in the following embodiments are as follows:

[0035] Calculation of wax reduction rate:

[0036] After the culture medium containing solid paraffin is degraded by bacteria in an incubator, it is filtered, centrifuged, air-dried, weighed and the mass of the remaining paraffin (M) is recorded. The paraffin degradation rate (V) is calculated using the following formula.

[0037] V = (M0 - M) × 100% / M0

[0038] Where: V—degradation rate of paraffin after reaction, %;

[0039] M0—Initial amount of unreacted paraffin, in g;

[0040] M — Mass of paraffin remaining after the reaction, in g.

[0041] Example 1: Optimization of the seed-liquid volume ratio of the compound bacteria

[0042] Slant culture: Alcanivorax dieselolei (CGMCC NO.1.8628), Alcanivorax dieselolei (CGMCC NO.1.3690), and Alcanivorax jadensis (CG MCC NO.1.16108) were inoculated into slant culture medium and cultured at 30℃ and 200rpm for 3 days.

[0043] Seed culture: Inoculate activated colonies into 20% seed culture medium and culture at 30°C and 200 rpm on a shaker until the final cell concentration reaches 10. 6 CFU / mL;

[0044] Seed culture preparation: Diesel-eating algae bacteria (CGMCC NO.1.8628) and Adriatic algae bacteria (CGMCC NO.1.16108) were mixed at viable cell ratios of 0:1, 1:2, 1:1, 2:1, and 1:0, while Diesel-eating algae bacteria (CGMCC No.1.3690) and Adriatic algae bacteria (CGMCC No.1.16108) were mixed at a viable cell ratio of 1:1 for fermentation culture.

[0045] Fermentation culture: The above-mixed seed liquid was inoculated into the fermentation medium at a rate of 6% v / v, and fermented for 168 h at a temperature of 30℃ and a shaking speed of 200 rpm.

[0046] The results are shown in Table 1 and Figure 1 As shown, under the same inoculum amount, the highest degradation rate of solid paraffin was achieved when the viable counts of Diesel-eating Alkyl Bacillus (CGMCC No. 1.8628) and Adriatic Alkyl Bacillus (CGMCC No. 1.16108) were mixed in a 1:1 ratio.

[0047] Table 1. Effects of different volume ratios of compound bacterial seed liquid on wax reduction rate

[0048]

[0049]

[0050] Preparation of inoculum:

[0051] (1) Slant culture: Alcanivorax dieselolei (CGMCC NO.1.8628) and Alcanivorax jadensis (CGMCC NO.1.16108) were inoculated into slant culture medium and cultured at 30℃ and 200rpm for 3 days.

[0052] (2) Seed culture: Activated colonies were inoculated into a 20% seed culture medium and cultured at 30℃ and 200 rpm on a shaker until the final cell concentration reached 1×10⁻⁶. 6 CFU / mL;

[0053] (3) Seed liquid compounding: Diesel edible algae bacteria (CGMCC NO.1.8628) and Adenomyces edible algae bacteria (CGMCC NO.1.16108) were mixed at a live bacteria ratio of 1:1.

[0054] (5) Freeze-drying: at a concentration of 1×10 6 Add 10% skim milk powder to the compound seed culture at CFU / mL, incubate at 37℃ for 1 hour, and then freeze dry to obtain the compound bacterial agent powder.

[0055] Example 2: Stability of the compound bacteria during passage

[0056] The optimal compounding ratio in Example 1 (i.e., when the viable counts of Diesel-eating Alkane Bacterium (CGMCC NO.1.8628) and Adenomyces yadense (CGMCC NO.1.16108) were compounded at a 1:1 ratio) was used for 12 generations of subculture. The strains after the 10th generation were inoculated into the fermentation medium for shake-flask fermentation, and the wax reduction rate after multiple subcultures was detected.

[0057] The results are shown in Table 2. The results indicate that the wax reduction rate still reached over 19.3% after 10 generations.

[0058] Table 2. Wax reduction rate after subculturing of compound bacteria

[0059]

[0060] Example 3: Optimization of Fermentation Medium

[0061] Under the optimal compound ratio conditions of Example 1, the fermentation medium conditions were optimized as follows:

[0062] 1. Optimization of carbon source for fermentation medium

[0063] The specific implementation steps are the same as in Example 1, except that the carbon source in the fermentation broth is glucose (1, 5, 10, 15, 20 g / L), and the other components in the culture medium are sodium nitrate 1.500 g / L, ammonium sulfate 1.500 g / L, dipotassium hydrogen phosphate 1.000 g / L, magnesium sulfate 0.500 g / L, potassium chloride 0.050 g / L, ferrous sulfate 0.010 g / L, and calcium chloride 0.002 g / L.

[0064] The results are shown in Table 3. When 10 g / L glucose was used as the carbon source, the wax reduction rate reached 26.4%. Therefore, 10 g / L glucose was selected as the carbon source.

[0065] Table 3. Dewaxing rate in fermentation media with different carbon source concentrations

[0066]

[0067] 2. Optimization of nitrogen source for fermentation medium

[0068] (1) Optimization of sodium nitrate

[0069] The specific implementation steps are the same as in Example 1, except that, based on the above optimization of the carbon source in the fermentation medium, the nitrogen source sodium nitrate in the fermentation medium is optimized.

[0070] The sodium nitrate concentrations were 1.000, 1.500, 2.000, 2.500, and 3.000 g / L, respectively. The other components in the culture medium were glucose 10 g / L, ammonium sulfate 1.500 g / L, dipotassium hydrogen phosphate 1.000 g / L, magnesium sulfate 0.500 g / L, potassium chloride 0.050 g / L, ferrous sulfate 0.010 g / L, and calcium chloride 0.002 g / L.

[0071] The results are shown in Table 4. When sodium nitrate of 2.000 g / L was used as the nitrogen source, the wax reduction rate reached 29.6%. Therefore, sodium nitrate of 2.000 g / L was selected.

[0072] Table 4. Dewaxing rate in fermentation media with different concentrations of sodium nitrate

[0073]

[0074] (2) Optimization of ammonium sulfate

[0075] The specific implementation steps are the same as in Example 1, except that the nitrogen source ammonium sulfate in the fermentation medium is optimized based on the optimal carbon source concentration and sodium nitrate concentration. The ammonium sulfate concentrations are 1.000, 1.500, 2.000, 2.500, and 3.000 g / L, respectively. The other components in the medium are glucose 10 g / L, sodium nitrate 2.000 g / L, dipotassium hydrogen phosphate 1.000 g / L, magnesium sulfate 0.500 g / L, potassium chloride 0.050 g / L, ferrous sulfate 0.010 g / L, and calcium chloride 0.002 g / L.

[0076] The results are shown in Table 5. When ammonium sulfate at 2.000 g / L was used as the nitrogen source, the wax reduction rate reached 31.7%. Therefore, ammonium sulfate at 2.000 g / L was selected.

[0077] Table 5. Dewaxing rate in fermentation media with different concentrations of ammonium sulfate

[0078]

[0079] (3) Optimization of phosphate in fermentation medium

[0080] The specific implementation steps are the same as in Example 1, except that, based on the above optimization of the optimal carbon source concentration, sodium nitrate concentration, and ammonium sulfate concentration, the phosphate in the fermentation medium is optimized.

[0081] The concentrations of dipotassium hydrogen phosphate were 0, 1.000, 2.000, 3.000, and 4.000 g / L, respectively. The other components in the culture medium were glucose 10 g / L, sodium nitrate 2.000 g / L, ammonium sulfate 2.000 g / L, magnesium sulfate 0.500 g / L, potassium chloride 0.050 g / L, ferrous sulfate 0.010 g / L, and calcium chloride 0.002 g / L.

[0082] The results are shown in Table 6. When the concentration of dipotassium hydrogen phosphate is 2.000 g / L, the wax reduction rate reaches 33.8%. Therefore, dipotassium hydrogen phosphate with a concentration of 2.000 g / L is selected.

[0083] Table 6. Dewaxing rate in fermentation media with different concentrations of dipotassium hydrogen phosphate

[0084]

[0085] (4) Optimization of metal ions in fermentation culture medium

[0086] The specific implementation steps are the same as in Example 1, except that the metal ions in the fermentation medium are optimized based on the optimal carbon source concentration, sodium nitrate concentration, ammonium sulfate concentration, and dipotassium hydrogen phosphate concentration.

[0087] ①Mg 2+ Optimization

[0088] The magnesium sulfate concentrations were 0, 0.500, 1.000, 1.500, and 2.000 g / L, respectively. The other components in the culture medium were glucose 10 g / L, sodium nitrate 2.000 g / L, ammonium sulfate 2.000 g / L, dipotassium hydrogen phosphate 2.000 g / L, potassium chloride 0.050 g / L, ferrous sulfate 0.010 g / L, and calcium chloride 0.002 g / L.

[0089] As shown in Table 7, the wax reduction rate reached 35.9% when the magnesium sulfate concentration was 0.700 g / L. Therefore, the magnesium sulfate concentration of 0.700 g / L was selected.

[0090] Table 7. Dewaxing rate in fermentation media with different concentrations of magnesium sulfate

[0091]

[0092]

[0093] ②Fe 2+ Optimization

[0094] The ferrous sulfate concentrations were 0, 0.010, 0.015, 0.020, and 0.025 g / L, respectively. Other components of the culture medium were: glucose 10 g / L, sodium nitrate 2.000 g / L, ammonium sulfate 2.000 g / L, dipotassium hydrogen phosphate 2.000 g / L, magnesium sulfate 0.700 g / L, potassium chloride 0.050 g / L, and calcium chloride 0.002 g / L. As shown in Table 8, the wax reduction rate reached 38.4% when the ferrous sulfate concentration was 0.015 g / L; therefore, a ferrous sulfate concentration of 0.015 g / L was selected.

[0095] Table 8. Dewaxing rate in fermentation media with different concentrations of ferrous sulfate

[0096] Concentration (g / L) Wax reduction rate 0 28.6％ 0.010 35.9％ 0.015 38.4％ 0.020 38.0％ 0.025 37.4％

[0097] ③Ca 2+ Optimization

[0098] The calcium chloride concentrations were 0, 0.002, 0.004, 0.006, and 0.008 g / L, respectively. Other components of the culture medium were: glucose 10 g / L, sodium nitrate 2.000 g / L, ammonium sulfate 2.000 g / L, dipotassium hydrogen phosphate 2.000 g / L, magnesium sulfate 0.700 g / L, potassium chloride 0.050 g / L, and ferrous sulfate 0.015 g / L. As shown in Table 9, the wax reduction rate reached 39.6% when the calcium chloride concentration was 0.004 g / L; therefore, a calcium chloride concentration of 0.004 g / L was selected.

[0099] Table 9. Dewaxing rate in fermentation media with different concentrations of calcium chloride

[0100] Concentration (g / L) Wax reduction rate 0 25.1％ 0.002 38.4％ 0.004 39.6％ 0.006 39.4％ 0.008 39.0％

[0101] Through optimization experiments, the optimal fermentation medium composition was finally determined to be:

[0102] Fermentation medium (g / L): glucose 10g / L, sodium nitrate 2.000g / L, ammonium sulfate 2.000g / L, dipotassium hydrogen phosphate 2.000g / L, magnesium sulfate 0.700g / L, potassium chloride 0.050g / L, ferrous sulfate 0.015g / L, calcium chloride 0.004g / L, solid paraffin 10g / L. After 168 hours of fermentation, the paraffin reduction rate reached 39.6%.

[0103] Example 4: Optimization of initial pH for shake flask fermentation

[0104] The conditions for slant culture and seed culture are the same as in Example 1.

[0105] Fermentation culture: The seed culture was inoculated into the fermentation medium at 6% v / v. The volume ratio of Diesel-eating Alkyl Bacillus (CGMC C NO.1.8628): Adenophora stricta (CGMCC NO.1.16108) in the compound bacterial seed culture was 1:1. The temperature was 30℃, the initial stirring speed was 200 rpm, and the initial pH was 6.0, 7.0, 8.0, 9.0, and 10.0.

[0106] The fermentation medium consisted of 10 g / L glucose, 2.000 g / L sodium nitrate, 2.000 g / L ammonium sulfate, 2.000 g / L dipotassium hydrogen phosphate, 0.700 g / L magnesium sulfate, 0.050 g / L potassium chloride, 0.015 g / L ferrous sulfate, and 0.004 g / L calcium chloride. Results are shown in Table 10. Figure 2 As shown, the wax reduction rate reached 43.2% when the initial pH value was 8.0, therefore the initial pH value was selected as 8.0.

[0107] Table 10 Effect of initial pH value on wax reduction rate

[0108] initial pH Wax reduction rate 6.0 35.7％ 7.0 39.6％ 8.0 43.2％ 9.0 40.7％ 10.0 36.4％

[0109] Example 5: Effect of solid paraffin concentration on fermentation

[0110] The conditions for slant culture, seed culture, and fermentation culture are the same as in Example 4.

[0111] Based on the above optimization of the fermentation medium and pH, the substance to be degraded (solid paraffin) was added, and further optimization was carried out by shaking flasks.

[0112] The concentrations of solid paraffin in the fermentation system were 10, 20, 30, 40, and 50 g / L, respectively. The fermentation medium consisted of 10 g / L glucose, 2.000 g / L sodium nitrate, 2.000 g / L ammonium sulfate, 2.000 g / L dipotassium hydrogen phosphate, 0.700 g / L magnesium sulfate, 0.050 g / L potassium chloride, 0.015 g / L ferrous sulfate, and 0.004 g / L calcium chloride. Solid paraffin was added at the initial stage of fermentation and continued until 168 hours of fermentation.

[0113] The results are shown in Table 11. When the concentration of added solid paraffin was 20 g / L, the paraffin reduction rate was similar to that of low-concentration paraffin, but more paraffin was degraded. The dynamic changes in the paraffin reduction rate in the shake flask under these conditions are shown in Table 11. Figure 3 As shown, the wax reduction rates reached 2.6%, 8.7%, 15.2%, 25.0%, 32.1%, 38.6%, and 43.0% at fermentation times of 24h, 48h, 72h, 96h, 120h, 144h, and 168h, respectively.

[0114] Table 11 Dewaxing rate in fermentation media with different concentrations of solid paraffin

[0115] Concentration (g / L) Wax reduction rate 10 43.2％ 20 43.0％ 30 28.5％ 40 20.2％ 50 15.4％

[0116] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. A microbial inoculant, characterized in that, The microbial agent contains Alcanivorax dieselolei (CGMCC No. 1.8628) and Alcanivorax jadensis (CGMCC No. 1.16108); The viable count ratio of Alcanivorax dieselolei (CGMCC No. 1.8628) to Alcanivorax jadensis (CGMCC No. 1.16108) was 1–2:1–2.

2. The microbial agent according to claim 1, characterized in that, The viable bacteria count in the microbial agent reaches 1 to 3 × 10⁻⁶. 6 CFU / g or CFU / mL.

3. The microbial agent according to claim 1, characterized in that, The microbial agent also contains the following components: glucose, sodium nitrate, ammonium sulfate, dipotassium hydrogen phosphate, magnesium sulfate, potassium chloride, ferrous sulfate, and calcium chloride.

4. The microbial agent according to claim 3, characterized in that, The concentrations of glucose, sodium nitrate, ammonium sulfate, dipotassium hydrogen phosphate, magnesium sulfate, potassium chloride, ferrous sulfate, and calcium chloride are 1–20 g / L, 1–3 g / L, 1–3 g / L, 1–4 g / L, 0–0.9 g / L, 0–0.08 g / L, 0–0.025 g / L, and 0–0.008 g / L, with a pH of 6–10.

5. The microbial agent according to claim 4, characterized in that, The concentrations of glucose, sodium nitrate, ammonium sulfate, dipotassium hydrogen phosphate, magnesium sulfate, potassium chloride, ferrous sulfate, calcium chloride, and pH were 10 g / L, 2 ...

6. The use of the microbial agent according to any one of claims 1 to 5 in the preparation of anti-wax preparations or degrading paraffin wax.

7. A method for degrading paraffin wax, characterized in that, The microbial agent described in any one of claims 1 to 5 is used to inoculate a solid paraffin culture medium for fermentation; wherein the concentration of solid paraffin is 10 to 50 g / L.

8. The method according to claim 7, characterized in that, The solid paraffin culture medium also contains: 1-20 g / L glucose, 1-3 g / L sodium nitrate, 1-3 g / L ammonium sulfate, 1-4 g / L dipotassium hydrogen phosphate, 0-0.9 g / L magnesium sulfate, 0-0.08 g / L potassium chloride, 0-0.025 g / L ferrous sulfate, and 0-0.008 g / L calcium chloride, with a pH of 6-10.

9. The method according to claim 7, characterized in that, The fermentation time is 168–192 hours and the fermentation temperature is 25–35℃.

10. The method according to claim 7, characterized in that, The concentration of the inoculant is 1–3 × 10⁻⁶. 6 CFU / mL.

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