Petrophilic strain and application thereof

Abstract

The invention relates to the field of microbial strains, in particular to a petroleum-philic strain and application thereof. The petroleum-philic strain is classified and named as Fusarium petroliphilum, the strain number of the petroleum-philic strain is GXGD-YZ-2513, the petroleum-philic strain is preserved in Guangdong Microbial Culture Collection Center on November 17, 2025, and the preservation number of the petroleum-philic strain is GDMCC No: 67305. The petroleum degrading bacterium has relatively strong stress resistance, and has obvious survival advantages and functional advantages in application in an actual environment. The strain has the capability of efficiently degrading petroleum, can tolerate the salt concentration of 0-10%, the temperature of 10-40 DEG C and the pH value of 3-11, and has the functional characteristic of tolerating high organic load. Meanwhile, the strain has a good degradation effect on vegetable oil and petroleum sewage. The invention provides a good strain and gene material for biological treatment of the environment polluted by crude oil.

Description

Technical Field

[0001] This invention relates to the field of microbial strains, specifically to a petroleum-loving strain and its applications. Background Technology

[0002] In recent years, petroleum has become one of the most important energy sources in modern society. During the exploration, extraction, refining, storage, transportation, and use of petroleum, accidents such as leaks can occur, causing serious environmental damage. Petroleum contains toxic alkanes (including alkanes and cycloalkanes), aromatic hydrocarbons (unsaturated hydrocarbons), and asphaltenes (sulfur-containing compounds and sulfur- or nitrogen-containing heterocyclic compounds). Some volatile organic compounds in petroleum hydrocarbons can affect human health, and some components have carcinogenic, mutagenic, and teratogenic effects. These compounds, with their pungent odors (benzene, xylene, etc.), are mainly chain compounds formed by the combination of carbon and hydrogen atoms, and they pose a significant hazard to water bodies.

[0003] In response to the increasingly serious problem of oil pollution, bioremediation methods utilize microorganisms to eliminate oil films on water surfaces and disperse dissolved petroleum hydrocarbons in water. It has significant advantages such as low cost, high efficiency, environmental friendliness, and no secondary pollution, and has become an effective method for solving oil pollution problems with both economic and environmental benefits.

[0004] Currently, researchers both domestically and internationally have screened and reported over 200 species of petroleum-degrading bacteria belonging to 70 genera. These include 28 genera of bacteria (such as *Pseudomonas*, *Acinetobacter*, and *Achromobacter*), 30 genera of molds (such as *Penicillium*, *Aspergillus*, and *Fusarium*), and 12 genera of yeasts (such as *Candida* and *Rhodotorula*). Among these, *Dietzia* sp., reported in Chinese patent "A Petroleum-Degrading Strains and Isolation Methods, Petroleum-Degrading Bacterial Agents and Their Preparation Methods and Applications" (CN 105505812 A), shows good petroleum degradation performance in neutral to slightly alkaline environments and can tolerate high salinity environments. The Chinese patent "A Petroleum-Degrading Bacterium KMJ-1 Producing Surfactants and Its Uses" (CN 116121111 A) reports that *Alcaligenes* sp. degrades oily sludge containing 5 g / L in scum. The sludge is placed in a shaking incubator at 150 r / min and 25 ℃ for 7 days, achieving a petroleum hydrocarbon degradation rate of up to 80.06%. The Chinese patent "A Petroleum-Degrading Bacterium with Phosphate-Solubilizing Activity and Its Cultivation Method and Application" (CN 111647528 A) reports that *Ochrobactrum daejeonense* possesses multiple functions including petroleum degradation, phosphate solubilization, and salt tolerance.

[0005] Currently, the degradation rate, degradation time, and stress resistance of petroleum-degrading bacteria still need further improvement. Therefore, exploring new and highly efficient petroleum-degrading bacteria and studying their degradation characteristics can provide excellent strains and genetic materials for the biological remediation of crude oil-polluted environments. Summary of the Invention

[0006] The technical problem to be solved by the first aspect of the present invention is to provide a petroleum-loving strain that addresses the shortcomings of existing technologies, such as low degradation rate, long degradation time, and poor stress resistance.

[0007] The technical problem to be solved by the second aspect of the present invention is to provide a degrading microbial agent.

[0008] The technical problem to be solved by the third aspect of the present invention is to provide the application of the petrophilic strain or degrading agent.

[0009] To solve the above-mentioned technical problems, the technical solution adopted by the present invention is as follows:

[0010] In the first aspect, the present invention provides a petrophilic strain, classified as Fusarium petroliphilum, strain number GXGD-YZ-2513, which was deposited on November 17, 2025 at the Guangdong Provincial Center for Microbial Culture Collection, with accession number GDMCC No: 67305, and the deposit address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0011] In some embodiments of the present invention, the petrophilic strain is a Fusarium, exhibiting aerobic characteristics. When cultured on PDA medium at 30°C for 3-5 days, the colony surface is dry and the colonies appear as white, fluffy particles. On potato dextrose agar, it shows concentric spreading characteristics, gradually turning yellow on the reverse side. This fungus is an asexual ascomycete, with large, sickle-shaped conidia. It reproduces through macroconidia, microconidia, and chlamydospores; the formation of chlamydospores enhances its survival ability and stress resistance.

[0012] In some embodiments of the present invention, the ITS characteristic sequence of the petrophilic strain is shown in SEQ ID NO.1 by molecular identification.

[0013] In some embodiments of the present invention, the petrophilic strain tolerates NaCl concentrations of 0-10%, temperatures of 10-40 °C, pH of 3-11, and petroleum loads of 0.2-22 g / L.

[0014] Secondly, the present invention provides a degrading bacterial agent containing the aforementioned petrophilic bacterial strain.

[0015] In some embodiments of the present invention, the degrading microbial agent is a petroleum-degrading microbial agent and / or an animal or vegetable oil-degrading microbial agent.

[0016] In some embodiments of the present invention, the degrading microbial agent is a petroleum-degrading microbial agent or a vegetable oil-degrading microbial agent.

[0017] In some embodiments of the present invention, the degrading agent is a solid degrading agent or a liquid degrading agent.

[0018] Thirdly, the present invention provides the application of the petrophilic strain in the degradation of oil pollutants.

[0019] In some embodiments of the present invention, the oil pollutants are petroleum pollutants or animal and vegetable oil pollutants, and the animal and vegetable oil pollutants are animal and vegetable oil wastewater.

[0020] In some embodiments of the present invention, the petroleum pollutants are petroleum hydrocarbon compounds and / or petroleum wastewater.

[0021] In some embodiments of the present invention, the petroleum contaminant is gasoline or gasoline wastewater.

[0022] In some embodiments of the present invention, the gasoline is 92# gasoline.

[0023] In some embodiments of the present invention, the animal and vegetable oil pollutants are vegetable oil wastewater.

[0024] In some embodiments of the present invention, the oil contaminants are emulsified by an emulsifier, which is any one of anhydrous ethanol, acetic acid, sodium carbonate, and sodium alkylbenzene sulfonate.

[0025] In some embodiments of the present invention, the oil contaminants are emulsified by an emulsifier, wherein the emulsifier is anhydrous ethanol.

[0026] Fourthly, the present invention provides the application of the aforementioned degrading microbial agent in the degradation of oil pollutants.

[0027] In some embodiments of the present invention, the oil pollutants are petroleum pollutants or animal and vegetable oil pollutants, and the animal and vegetable oil pollutants are animal and vegetable oil wastewater.

[0028] In some embodiments of the present invention, the petroleum pollutants are petroleum hydrocarbon compounds and / or petroleum wastewater.

[0029] In some embodiments of the present invention, the petroleum contaminant is gasoline or gasoline wastewater.

[0030] In some embodiments of the present invention, the gasoline is 92# gasoline.

[0031] In some embodiments of the present invention, the animal and vegetable oil pollutants are vegetable oil wastewater.

[0032] In some embodiments of the present invention, the oil contaminants are emulsified by an emulsifier, which is any one of anhydrous ethanol, acetic acid, sodium carbonate, and sodium alkylbenzene sulfonate.

[0033] In some embodiments of the present invention, the oil contaminants are emulsified by an emulsifier, wherein the emulsifier is anhydrous ethanol.

[0034] In some embodiments of the present invention, the petrophilic strain can degrade petroleum wastewater containing different concentrations of petroleum within 28 days, with a degradation rate of 84.21-90.58%.

[0035] In some embodiments of the present invention, the petrophilic strain can degrade vegetable oil wastewater containing different concentrations of vegetable oil within 28 days, with a degradation rate of 90.17-99.04%.

[0036] Beneficial effects:

[0037] This invention screened a petrophilic bacterial strain from petroleum-contaminated soil samples, named *Fusarium petroliphilum*, strain number GXGD-YZ-2513, and deposited at the Guangdong Provincial Microbial Culture Collection Center on November 17, 2025, with accession number GDMCC No: 67305. This petrophilic degrading bacterium exhibits strong stress resistance, demonstrating significant survival and functional advantages in practical applications. This strain possesses highly efficient petroleum degradation capabilities, tolerating salt concentrations of 0–10%, temperatures of 10–40 °C, and pH values ​​of 3–11, and also exhibits tolerance to high organic loads. Furthermore, this strain demonstrates good degradation effects on both vegetable oils and petroleum wastewater, with degradation rates reaching 84.21–99.05%. This invention provides a valuable bacterial strain and genetic material for the biological remediation of crude oil-contaminated environments. Attached Figure Description

[0038] The present invention will be further described in detail below with reference to the accompanying drawings, and the advantages of the present invention in the above and / or other aspects will become clearer.

[0039] Figure 1 This is a diagram showing the disappearance of the oil layer during the enrichment and domestication culture of the strains in this embodiment of the invention.

[0040] Figure 2 This is a colony diagram of the petrophilic strain GXGD-YZ-2513 in an embodiment of the present invention.

[0041] Figure 3 This is a microscopic colony image of the petrophilic strain GXGD-YZ-2513 in an embodiment of the present invention.

[0042] Figure 4Images of the petrophilic strain GXGD-YZ-2513 grown under different salinity conditions for 4 days in an embodiment of the present invention.

[0043] Figure 5 Images of the petrophilic strain GXGD-YZ-2513 grown under different pH conditions for 4 days in this embodiment of the invention.

[0044] Figure 6 Images of the petrophilic strain GXGD-YZ-2513 grown for 4 days under different temperature conditions in this embodiment of the invention.

[0045] Figure 7 Images of the petrophilic strain GXGD-YZ-2513 grown for 4 days under different petroleum concentrations in this embodiment of the invention.

[0046] Figure 8 The results of selecting petroleum or vegetable oil emulsifying solvents in the embodiments of the present invention.

[0047] Figure 9 This illustrates the degradation of vegetable oil in an embodiment of the present invention.

[0048] Figure 10 This illustrates the degradation of petroleum in an embodiment of the present invention. Detailed Implementation

[0049] The present invention will be further described in detail below with reference to specific embodiments, and the advantages of the present invention in the above and / or other aspects will become clearer.

[0050] Unless otherwise specified, the experimental methods described in the following examples are conventional methods; the reagents and materials described are commercially available unless otherwise specified.

[0051] In the following embodiments, the petroleum is Sinopec 92# gasoline.

[0052] In the following examples, the inorganic salt culture (MSM) consisted of: 1.0 g / L NH4Cl, 0.06 g / L CaCl2, 0.15 g / L MgSO4·7H2O, 1.0 g / L K2HPO4, 1.0 g / L KH2PO4, and pH 7.0.

[0053] The petroleum liquid culture medium (SY) consists of: 1.0 g / L NH4Cl, 0.06 g / L CaCl2, 0.15 g / L MgSO4·7H2O, 1.0 g / L K2HPO4, 1.0 g / L KH2PO4, pH 7.0, and petroleum suspension at a concentration of 0.2–22 g / L. The petroleum suspension is prepared by adding petroleum to a solvent at a 1:1 volume / mass ratio. The solvent can be any one of anhydrous ethanol, acetic acid, sodium carbonate, and sodium alkylbenzene sulfonate.

[0054] The solid separation culture medium (GSY) consists of: 1.0 g / L NH4Cl, 0.06 g / L CaCl2, 0.15 g / L MgSO4·7H2O, 1.0 g / L K2HPO4, 1.0 g / L KH2PO4, pH 7.0, 20 g / L petroleum suspension, and 20 g / L agar.

[0055] The PDA solid culture medium consists of: 200 g / L potato juice, 20 g / L glucose, 3 g / L potassium dihydrogen phosphate, 1.5 g / L magnesium sulfate, and 0.1 g / L vitamin B1.

[0056] The liquid culture medium for vegetable oil consists of: 1.0 g / L NH4Cl, 0.06 g / L CaCl2, 0.15 g / L MgSO4·7H2O, 1.0 g / L K2HPO4, 1.0 g / L KH2PO4, and 2–22 g / L vegetable oil suspension, pH 7.0. The vegetable oil suspension is prepared by adding petroleum to a solvent at a 1:1 volume / mass ratio. The solvent can be any one of anhydrous ethanol, acetic acid, sodium carbonate, and sodium alkylbenzene sulfonate.

[0057] Example 1: Isolation and purification of petrophilic strains

[0058] (1) Take petroleum-contaminated soil samples from Xiaohuangni River in Panzhou City, Liupanshui City, Guizhou Province, add inorganic salt culture medium (MSM), shake to fully disperse the soil, and let it settle.

[0059] (2) Add the supernatant obtained by static precipitation in step (1) to petroleum liquid culture medium (SY) containing 20 g / L petroleum suspension, and culture in a shaker at 30 ℃ and 200 rpm.

[0060] (3) Select the bottle of bacterial culture with the turbid liquid, the disappearance of the oil layer or the thinnest layer and transfer it to fresh petroleum liquid culture medium containing 20 g / L petroleum suspension at a ratio of 20% v / v. Culture at 30 °C and 200 rpm until the liquid becomes turbid and the oil layer disappears. Then, transfer it to enrichment culture at a ratio of 10%, 5%, 2% and 1% v / v in sequence. Culture a total of 5 batches to obtain enriched bacterial culture. Figure 1 This is a diagram showing the disappearance of the oil layer during the enrichment and domestication culture of the strains in this embodiment of the invention.

[0061] (4) Dilute the enriched bacterial culture obtained in step (3) to 10. -3 10 -5 10 -7 Then, 100 μL of the bacterial suspension was spread onto fresh solid isolation medium (GSY) and incubated at 30 °C for 3-5 days. Colonies with good growth and clear oleophilic zones were selected, streaked, and re-inoculated into petroleum liquid medium (SY). The mixture was purified three times, spread onto PDA solid medium, and the bacterial cells were collected to obtain the petroleum-loving strain.

[0062] Example 2: Identification of petrophilic strains

[0063] 1. Morphological identification

[0064] Morphological identification of petrophilic strains. Figure 2 This is a colony diagram of petrophilic strains in the culture medium, created by... Figure 2 As can be seen, the colony surface is dry, and the colony is white and fluffy. It exhibits concentric circle spreading characteristics on potato dextrose agar medium, and the reverse side gradually turns yellow. This strain is aerobic, and after being cultured on PDA medium at 30 ℃ for 3-5 days, the diameter of the bacteria reaches 60-90 cm.

[0065] 2. Identification of microstructure morphology

[0066] The microstructure and morphology of petrophilic strains were identified. Figure 3 These are microscopic images of colonies of petrophilic bacterial strains, produced by [Author Name]. Figure 3 It can be seen that the microstructure of this strain shows a large number of macroconidia, microconidia and chlamydospores, which are elliptical, 6-10 μm long and 2-3 μm wide; the hyphae are septate, branched, 2-8 μm wide and more than 100 μm long.

[0067] 3. Molecular level identification

[0068] The molecular genetic classification of the petrophilic strain was identified using ITS1 / ITS4 sequencing and analysis at the molecular level. PCR amplification was performed using universal fungal primers: ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') for the forward primer and ITS4 (5'-TCCTCCGCTTATTGATATGC-3') for the reverse primer. The amplified products were sent to Beijing Ruiboxing Technology Co., Ltd., where they were electrophoresed, purified, and their characteristic ITS sequence was determined, as shown in SEQ ID NO.1. This characteristic sequence was compared for homology with existing nucleic acid sequences in the NCBI gene bank. The strain showed 99.13% identity with *Fusarium petroliphilum*. Therefore, based on morphological and structural characteristics, this strain can be identified as *Fusarium petroliphilum*.

[0069] The petrophilic strain was preserved in the company's self-built library, "Guangdong University of Technology Biotechnology Research Institute Co., Ltd. Microbial Library," and obtained the internal number GXGD-YZ-2513. On November 17, 2025, the strain was deposited at the Guangdong Provincial Microbial Culture Collection Center, classified and named Fusarium petroliphilum, with the strain number GXGD-YZ-2513, the accession number GDMCCNo: 67305, and the deposit address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0070] Example 3: Salt tolerance of petrophilic strain GXGD-YZ-2513

[0071] (1) Plate activation: Under sterile conditions, the frozen petrophilic strain GXGD-YZ-2513 was streaked onto PDA solid medium and cultured at 30 ℃ for 1~2 days.

[0072] (2) The activated strains obtained in step (1) were inoculated into PDA medium containing salt at different concentration gradients, and their growth was observed and measured on day 4. The salt concentrations in the PDA medium were 1%, 2%, 3%, 5%, 7%, and 10% of NaCl by mass fraction, respectively.

[0073] Table 1 shows the growth of the petrophilic strain GXGD-YZ-2513 under different salinity conditions. Figure 4 Images of the petrophilic strain GXGD-YZ-2513 after 4 days of growth under different salinity conditions. (From Table 1 and...) Figure 4 It can be seen that the salinity tolerance range of the petroleum-loving strain GXGD-YZ-2513 is 0~10%, with an optimal salinity of 1%, and it can still grow when the salinity reaches above 7%.

[0074] Table 1. Growth of petrophilic strain GXGD-YZ-2513 under different salinity conditions.

[0075]

[0076] Example 4: pH tolerance of petrophilic strain GXGD-YZ-2513

[0077] (1) Plate activation: Under sterile conditions, the frozen petrophilic strain GXGD-YZ-2513 was streaked onto PDA solid medium and cultured at 30 ℃ for 1~2 days.

[0078] (2) The activated strains obtained in step (1) were inoculated into PDA medium with different pH values, and their growth was observed and measured on day 4. The pH values ​​of the PDA medium were controlled at 3, 5, 7, 9 and 11, respectively.

[0079] Table 2 shows the growth of the petrophilic strain GXGD-YZ-2513 under different pH conditions. Figure 5 Images of the petrophilic strain GXGD-YZ-2513 grown under different pH conditions for 4 days, as shown in Table 2 and... Figure 5 It can be seen that the petrophilic strain GXGD-YZ-2513 has a relatively wide pH tolerance range, especially favoring growth under alkaline conditions, with a pH tolerance range of 3-11. When the pH is 3, the plate fails to solidify; this can be confirmed by measuring the OD... 600 Judging from the growth of the strain, it can be found that although the strain does not grow as well as under other pH conditions at pH 3, it can still grow.

[0080] Table 2. Growth of petrophilic strain GXGD-YZ-2513 under different pH conditions

[0081]

[0082] Example 5: Temperature tolerance of petrophilic strain GXGD-YZ-2513

[0083] (1) Plate activation: Under sterile conditions, the frozen petrophilic strain GXGD-YZ-2513 was streaked onto PDA solid medium and cultured at 30 ℃ for 1~2 days.

[0084] (2) The activated strain obtained in step (1) was inoculated into PDA medium and cultured under different temperature conditions. The growth on day 4 was observed and measured. The culture temperature was controlled at 10 ℃, 20 ℃, 30 ℃ and 40 ℃ respectively.

[0085] Table 3 shows the growth of the petrophilic strain GXGD-YZ-2513 under different temperature conditions. Figure 6 Images of the petrophilic strain GXGD-YZ-2513 grown for 4 days under different temperature conditions, as shown in Table 3 and... Figure 6 It can be seen that strain GXGD-YZ-2513 can tolerate temperatures of 10~40 ℃, with the optimal temperature being 30 ℃. The high temperature of 40 ℃ showed inhibition, but it could still grow.

[0086] Table 3. Growth of petrophilic strain GXGD-YZ-2513 under different temperature conditions.

[0087]

[0088] Example 6: Tolerance of petroleum-loving strain GXGD-YZ-2513 to petroleum

[0089] (1) Plate activation: Under sterile conditions, the frozen petrophilic strain GXGD-YZ-2513 was streaked onto PDA solid medium and cultured at 30 ℃ for 1~2 days.

[0090] (2) The activated strains obtained in step (1) were inoculated into petroleum liquid culture medium containing different concentrations of petroleum suspension (1 g / L, 2 g / L, 3 g / L, 5 g / L, 10 g / L, 20 g / L), and their growth on day 4 was observed and measured.

[0091] Table 4 shows the growth of the petrophilic strain GXGD-YZ-2513 under different petroleum concentrations. Figure 7 Images of the petrophilic strain GXGD-YZ-2513 after 4 days of growth under different petroleum concentrations, as shown in Table 4. Figure 7 It can be seen that the growth of petroleum-loving strain GXGD-YZ-2513 is not significantly affected by the concentration of petroleum suspension at 1~10 g / L; when the concentration reaches 20 g / L, it can still maintain a growth state of 26.5±0.707 mm.

[0092] Table 4. Growth of strain GXGD-YZ-2513 under different petroleum suspension concentrations.

[0093]

[0094] Example 7: Selection of petroleum or vegetable oil emulsifiers

[0095] Add petroleum or vegetable oil to the emulsifier at a 1:1 (volume / mass ratio), wherein the emulsifier is any one of anhydrous ethanol, acetic acid, sodium carbonate, and sodium alkylbenzene sulfonate. Shake well until homogeneous, and let stand overnight. Figure 8 .

[0096] according to Figure 8 Results analysis showed a comparison of results before and after adding emulsifiers. Before adding emulsifiers, oil and water separation occurred. After adding emulsifiers, the emulsification effect was obvious. Among them, the addition of anhydrous ethanol had the best effect. Anhydrous ethanol will be used as emulsifier in the subsequent experiments.

[0097] Example 8: Evaluation of the degradation of vegetable oil by petrophilic strain GXGD-YZ-2513

[0098] (1) Preparation of seed culture: Under sterile conditions, the isolated and purified petrophilic strain GXGD-YZ-2513 was picked and inoculated into a 500 mL shake flask containing 100 mL PDA liquid culture medium and cultured at 30 ℃ and 200 rpm for 48 h.

[0099] (2) Seed culture of petrophilic strain GXGD-YZ-2513 was inoculated at 10% v / v into liquid culture medium containing different concentrations of plant oil and cultured at 30 ℃ and 200 rpm for 28 days. The growth of the petrophilic strain and the degradation of plant oil were observed and measured every 3-4 days. The formula for calculating the degradation rate of plant oil is as follows:

[0100]

[0101] (3) The oil content in the solution was determined by using an infrared oil analyzer (equipment model JLBG-12N series infrared spectrophotometer, Jilin Jiguang Technology Co., Ltd.).

[0102] Using the petrophilic strain GXGD-YZ-2513 without the strain as a blank group, the degradation of vegetable oil was observed at initial concentrations of (20807.22±544.94) mg / L and (2090.76±0.91) mg / L, respectively, with the presence of the petrophilic strain. The results are shown in Table 5. Figure 9As shown in the figure. In the blank group, the volatility of vegetable oil was controlled to below 15% by adding anhydrous ethanol. In vegetable oil at a concentration of (20807.22±544.94) mg / L, the degradation of vegetable oil reached 41.76% on day 10, 63.92% on day 13, 85.20% on day 16, and 90.03-90.17% on days 25-28. Degradation stopped when the vegetable oil concentration was controlled at around 2000 mg / L. In vegetable oil with a concentration of (2090.76±0.91) mg / L, the degradation of vegetable oil can reach 72.66% on day 10, 84.60% on day 13, 95.96% on day 16, and 99.04-99.05% on days 25-28. The oil concentration can be controlled below 20 mg / L, which can meet the national standard GB 31571-2015 indirect emission standard.

[0103] The above results indicate that the petrophilic strain GXGD-YZ-2513 has a significant ability to degrade vegetable oils, especially at low concentrations, achieving efficient and near-complete degradation, and possesses good application potential. Based on the degradation results at both high and low concentrations, this strain can be further enhanced by employing a secondary concentration-enhancing strategy to further increase degradation efficiency at high concentrations of vegetable oils.

[0104] Table 5. Degradation of vegetable oils by petrophilic strain GXGD-YZ-2513

[0105]

[0106] Example 9: Evaluation of petroleum degradation by petroleum-loving strain GXGD-YZ-2513

[0107] (1) Preparation of seed culture: Under sterile conditions, the isolated and purified petrophilic strain GXGD-YZ-2513 was picked and inoculated into a 500 mL shake flask containing 100 mL PDA liquid culture medium and cultured at 30 ℃ and 200 rpm for 48 h.

[0108] (2) The petrophilic strain GXGD-YZ-2513 was inoculated into petroleum liquid culture medium containing different concentrations of petroleum and cultured at 30 ℃ and 200 rpm for 28 days. The growth of the petrophilic strain and the petroleum degradation were observed and measured every 3-4 days. The formula for calculating the petroleum degradation rate is as follows:

[0109]

[0110] (3) The oil content in the solution was determined by using an infrared oil analyzer (equipment model JLBG-12N series infrared spectrophotometer, Jilin Jiguang Technology Co., Ltd.).

[0111] Using the petrophilic strain GXGD-YZ-2513 as the blank group, Table 6 and Figure 10 The degradation of petroleum by the petroleum-loving strain GXGD-YZ-2513 at different concentrations (20807.22±544.94) mg / L, (2087.22±121.94) mg / L, and (202.79±0.42) mg / L was demonstrated. In the control group, the volatility of petroleum was controlled to below 7% by adding anhydrous ethanol. The petrophilic strain GXGD-YZ-2513 exhibited the following degradation rates for petroleum: At an initial petroleum concentration of (20807.22±544.94) mg / L, the degradation rate reached 25.38% on day 10, 51.40% on day 15, 67.80% on day 21, and 84.21% on day 28, at which point the petroleum concentration could be controlled at approximately 3300 mg / L. At an initial petroleum concentration of (2087.22±121.94) mg / L, the degradation rate reached 44.58% on day 10, 65.83% on day 15, 85.17% on day 21, and 87.83% on day 28, at which point the petroleum concentration could be controlled at approximately 250 mg / L. The initial petroleum concentration was approximately 202.79 ± 0.42 mg / L. The degradation rate reached 51.78% on day 10, 67.27% on day 15, 82.44% on day 21, and 90.58% on day 28. At this point, the petroleum concentration could be controlled below 20 mg / L, meeting the national standard GB31571-2015 for indirect emissions.

[0112] The above results indicate that the petrophilic strain GXGD-YZ-2513 also has a significant ability to degrade petroleum.

[0113] Table 6. Degradation of petroleum by petrophilic strain GXGD-YZ-2513

[0114]

[0115] This invention provides a petroleum-loving bacterial strain and its applications, along with related ideas and methods. Many methods and approaches exist for implementing this technical solution; the above description is merely a preferred embodiment of the invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of this invention, and these improvements and modifications should also be considered within the scope of protection of this invention. All components not explicitly stated in this embodiment can be implemented using existing technologies.

Claims

1. A petrophilic strain, classified as Fusarium petroliphilum, strain number GXGD-YZ-2513, was deposited at the Guangdong Provincial Microbial Culture Collection Center on November 17, 2025, with accession number GDMCC No: 67305.

2. The petrophilic strain according to claim 1, characterized in that, The ITS characteristic sequence of the petrophilic strain is shown in SEQ ID NO.

1.

3. The petrophilic strain according to claim 1, characterized in that, The petroleum-loving strains tolerated NaCl concentrations of 0–10%, temperatures of 10–40 °C, pH values ​​of 3–11, and petroleum loads of 0.2–22 g / L.

4. A biodegrading agent, characterized in that, The degrading agent contains the petrophilic strain described in claim 1.

5. The degrading bacterial agent according to claim 4, characterized in that, The degrading agent is a petroleum-degrading agent and / or an animal or vegetable oil-degrading agent.

6. The degrading microbial agent according to claim 4 or 5, characterized in that, The degrading agent is either a solid degrading agent or a liquid degrading agent.

7. The application of the petrophilic strain according to any one of claims 1 to 3 in the degradation of oil pollutants; wherein, The oil pollutants are petroleum pollutants or animal and vegetable oil pollutants.

8. The application according to claim 7, characterized in that, The petroleum pollutants are petroleum hydrocarbon compounds and / or petroleum wastewater; the animal and vegetable oil pollutants are animal and vegetable oil wastewater.

9. The application of the degrading microbial agent according to any one of claims 4 to 6 in the degradation of oil pollutants; wherein, The oil pollutants are petroleum pollutants or animal and vegetable oil pollutants.

10. The application according to claim 9, characterized in that, The petroleum pollutants are petroleum hydrocarbon compounds and / or petroleum wastewater; the animal and vegetable oil pollutants are animal and vegetable oil wastewater.

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