Microbial communities with soil remediation functions and their applications

Abstract

This invention provides a microbial community with petroleum-degrading capabilities, comprising *Gordonia sp. Strain C1*, *Cellulosimicrobium sp. Strain C3*, or both, wherein *Gordonia sp. Strain C1* and *Cellulosimicrobium sp. Strain C3* are CGMCC No. 25708 and CGMCC No. 25709, respectively. Using the microbial community and culture formula provided by this invention, the community achieves a petroleum hydrocarbon degradation rate greater than 60% in a petroleum degradation system with a petroleum content of 60,000-100,000 mg / L, with a maximum degradation rate exceeding 85%. This community can be applied in petroleum pollution treatment processes centered on microorganisms, contributing to the ecological remediation of petroleum-polluted environments and showing broad application prospects in the ecological restoration of petroleum-polluted soil or water bodies.

Description

Technical Field

[0001] This invention relates to the field of environmental biotechnology, specifically to microbial communities with petroleum degradation functions and their applications. Background Technology

[0002] Petroleum is a vital energy source for social development, and the petroleum industry plays an irreplaceable role in driving global economic growth and improving social life. However, oil pollution accidents occur during extraction, transportation, use, and storage, and the frequency of oil spills is showing a significant upward trend. Because petroleum contains a highly complex array of compounds, including many highly toxic components that are difficult to degrade and easily accumulate, oil spills pose a serious threat to environmental safety and, consequently, human health. With the gradual improvement of societal standards and the increasing awareness of environmental protection, how to address oil spills and the resulting environmental pollution has become a major global concern.

[0003] Petroleum is a mixture of various organic compounds with highly diverse chemical compositions. In oil-polluted environments, it exhibits uneven dispersion and high heterogeneity, posing significant challenges to the effective treatment of such pollution. Over the past few decades, petroleum pollution treatment processes based on physical, chemical, and biological methods have been developed. Among these, bioremediation technologies, utilizing microorganisms to biodegrade petroleum-polluted environments, have attracted considerable attention due to their environmental friendliness, high efficiency, and low cost compared to other strategies for treating petroleum pollution.

[0004] Currently, domestic research on petroleum-degrading microorganisms is actively underway for the microbial remediation of petroleum-polluted environments. Existing studies largely focus on single petroleum-degrading microorganisms, and the culture conditions are not optimized based on strain characteristics, as exemplified by patents CN111718867B, CN105505812B, and CN109777747A. Furthermore, due to the limited degradation efficiency of single-species degrading bacteria, mixed bacterial degradation systems can achieve efficient and rapid degradation of petroleum pollutants, and their construction has attracted considerable attention, as seen in patents CN110511890A and CN111534462A. However, most mixed systems suffer from low petroleum hydrocarbon degradation efficiency or low degradation concentrations. Therefore, there is an urgent need to develop a composite microbial community with petroleum-degrading capabilities, capable of efficiently degrading high concentrations of petroleum pollutants. Summary of the Invention

[0005] In view of this, the main objective of the present invention is to provide a composite microbial community with petroleum degradation function and its application, in order to solve the aforementioned technical problems.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0007] The two petroleum-degrading functional bacteria provided by this invention were isolated from oil sludge in the No. 1 Oil Production Plant of Qinghai Oilfield through single-strain screening. The two obtained petroleum-degrading functional bacteria were subjected to colony morphology and molecular biological identification. Identification showed that the obtained C1 colony was orange-red, opaque, and raised; the C3 colony changed from white to green, was opaque, and raised. Based on the maximum likelihood method and a phylogenetic tree constructed using MEGA 7.0 software, the strains were sequentially named *Gordonia sp.*, named *Gordonia sp.Strain C1*, and *Cellulosimicrobium sp.*, named *Cellulosimicrobium sp.Strain C3*.

[0008] The obtained single petroleum-degrading bacteria, Gordonia sp. Strain C1 and Cellulosimicrobium sp. Strain C3, were subjected to a unique carbon source utilization test.

[0009] This invention provides culture formulations and conditions for two petroleum-degrading functional bacteria. Culture formulation: BO medium; Culture conditions: 26℃-37℃, 150-200 rpm.

[0010] As another aspect of the present invention, the application conditions of the compound bacterial solution with petroleum degradation function are provided, wherein the culture medium uses a modified BO culture formula and the culture conditions are 26℃-37℃ and 150-200rpm.

[0011] This invention also provides a method for culturing and combining two petroleum-degrading functional bacteria, wherein *Gordonella foenum-graecum* C1 and *Microbes fibrocarpa* C3 are cultured separately; and the concentrations (OD) of the two bacterial cultures are adjusted. 600 The concentration is 0.6-1.0, and the mixture is simultaneously mixed and compounded under the conditions of 50%-70% and 30%-50% respectively.

[0012] This invention also provides the application of the aforementioned microbial community in degrading petroleum hydrocarbons in petroleum-polluted environments.

[0013] The present invention also provides formulations for the aforementioned microbial community in petroleum degradation systems.

[0014] The present invention also provides a method for degrading petroleum hydrocarbons. A mixed bacterial community is inoculated at a rate of 10%-20% into a degradation system with a petroleum mass concentration of 60,000-100,000 mg / L, and cultured at 26℃-37℃ and 150-200 rpm for 8-24 days to achieve petroleum degradation.

[0015] More specifically, the present invention includes the following:

[0016] 1. A bacterial community, characterized in that the bacterial community comprises Gordonia sp. C1 with accession number CGMCC No. 25708, Cellulosimicrobium sp. C3 with accession number CGMCC No. 25709, or both.

[0017] 2. The microbial community according to claim 1, characterized in that the microbial community consists of 50%-70% by volume of Gordonia sp. C1 and 30%-50% by volume of Cellulosimicrobium sp. C3.

[0018] 3. A method for culturing the microbial community described in item 1 or 2, characterized in that the culture medium used for culturing has the following composition: CH3COONa 1g / L, C3H3NaO3 0.5g / L, K2HPO4 0.5g / L, KNO3 0.1g / L, NH4Cl 0.05g / L, NaCl 25g / L, FeC6H5O7 0.002g / L, natural pH; and the culture conditions are: 26℃-37℃, 150-200rpm.

[0019] 4. The application of the microbial community described in item 1 or 2 in the field of oil pollution environmental remediation.

[0020] 5. According to the application described in item 4, the oil-polluted environment includes oil-polluted soil and oil-polluted water.

[0021] 6. A method for microecological remediation of petroleum pollution, characterized in that the method uses the microbial community described in item 1 or 2 to degrade petroleum hydrocarbons in petroleum-polluted soil or water.

[0022] 7. A method for degrading petroleum hydrocarbons, characterized in that the method involves inoculating the bacterial community of claim 1 or 2 into a degradation system containing petroleum at a mass concentration of 60,000-100,000 mg / L and culturing it at 26℃-37℃ and 150-200 rpm.

[0023] 8. The method according to item 7, characterized in that the nutrients in the degradation system are CH3COONa 0.3g / L, C3H3NaO3 0.1g / L, K2HPO4 1g / L, KNO3 2g / L, NH4Cl 0.5g / L, NaCl 10g / L, and FeC6H5O7 0.005g / L.

[0024] 9. An article for treating petroleum-polluted environments, characterized in that the article contains the microbial community described in item 1 or 2.

[0025] 10. The article according to item 9, characterized in that the article is a liquid microbial agent article.

[0026] Compared with the prior art, the present invention has the following beneficial technical effects:

[0027] The microbial community with petroleum degradation function provided by this invention can achieve efficient degradation of high concentrations of petroleum pollutants (60,000-100,000 mg / L), and the degradation efficiency is significantly improved when two strains are used together compared with that of a single strain. Attached Figure Description

[0028] Figure 1 This is a photograph of a culture of Gordonia sp. Strain C1.

[0029] Figure 2 This is a photograph of the culture of Cellulosimicrobium sp. Strain C3.

[0030] Figure 3 The growth curves of the strain at 37℃ under different culture medium conditions are shown.

[0031] Figure 4 The growth curves of the strain under different temperature conditions in BO medium are shown.

[0032] Figure 5 The graph shows the degradation rate of petroleum hydrocarbons by the strain after 24 days in a degradation system with a petroleum mass concentration of 80,000 mg / L.

[0033] Figure 6 The C1 of the strain after 24 days of degradation in a petroleum degradation system with a concentration of 80,000 mg / L. 10 -C 40 GC-MS peak diagram of aliphatic hydrocarbons;

[0034] Figure 7 The graph shows the degradation rate of petroleum hydrocarbons after 8, 16, and 24 days of biodegradation of the composite bacterial solution in a degradation system with a petroleum mass concentration of 80,000 mg / L.

[0035] Information on the preservation of biological material samples:

[0036] Gordonia sp. strain C1, taxonomically named Gordonia sp., was deposited on September 15, 2022, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with accession number CGMCCNo. 25708.

[0037] Cellulosimicrobium sp. strain C3, classified as Cellulosimicrobium sp., was deposited on September 15, 2022, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, with accession number CGMCC No. 25709. Detailed Implementation

[0038] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments and accompanying drawings. These embodiments are merely illustrative examples and are not intended to limit the invention. Unless otherwise specified, the experimental methods in the following embodiments are standard experimental procedures. All instruments, consumables, and reagents used in the following embodiments are commercially available unless otherwise specified. In the quantitative experiments in the following embodiments, three biological replicates are set up for each experiment.

[0039] The culture medium formulation used in the following examples is as follows:

[0040] Microbial enrichment medium: 20mM NH4Cl, 5mM K2HPO4, natural pH.

[0041] LB1 medium: NaCl 15g / L, yeast 3g / L, tryptone 7g / L, natural pH.

[0042] Biodegradable (BO) medium: CH3COONa 1g / L, C3H3NaO3 0.5g / L, K2HPO4 0.5g / L, KNO3 0.1g / L, NH4Cl 0.05g / L, NaCl 25g / L, FeC6H5O7 0.002g / L, natural pH. Sterilize at 115℃ for 30min.

[0043] Modified BO medium: CH3COONa 0.3 g / L, C3H3NaO3 0.1 g / L, K2HPO4 1 g / L, KNO3 2 g / L, NH4Cl 0.5 g / L, NaCl 10 g / L, FeC6H5O7 0.005 g / L, natural pH. Sterilize at 115℃ for 30 min.

[0044] Petroleum degradation medium: Add 80,000 mg / L of petroleum to BO medium and sterilize at 115°C for 45 min.

[0045] Petroleum screening plate medium: Based on the microbial enrichment medium, add Tween 80 2.5 mL / L, sodium hexametaphosphate 0.2 g / L, polyvinylpyrrolidone K30 0.05 g / L, 10% SDS 0.5 mL / L, 0.5 M pH=8.0 EDTA 1 mL / L, petroleum 100000 mg / L, and agar 15 g / L in sequence. After thorough mixing, sterilize at 115℃ for 45 min.

[0046] Gas chromatography-mass spectrometry (GC-MS-QP2010 Ultra, Japan), multi-functional microplate reader (molecular devices SpectraMax i3x, USA).

[0047] Example 1: Isolation, identification and carbon source utilization test of petroleum-degrading functional bacteria

[0048] Approximately 30g of crude oil-contaminated soil was added to 200mL of microbial enrichment medium. Sterilized petroleum (5-10g) and 5-20mL of microbial enrichment medium were added every 30 days to construct a 200mL petroleum-degrading microbial enrichment reactor. After 180 days of enrichment and acclimatization culture following the construction of the reactor, the microorganisms were transferred at a 20% inoculum to an enrichment medium containing 15% crude oil (m / v). Six cycles of enrichment and selection culture were then conducted, with each cycle lasting 15-20 days. The temperature was maintained at 30℃-37℃ and the rpm was 150-180.

[0049] The enriched bacterial culture was serially diluted to 10 μL using 1×PBS. -6 10 -7 and 10 -8 Single colonies were streaked onto petroleum screening plates using petroleum as the sole carbon source and incubated at 37°C for 2-4 days. Colonies of different sizes, colors, and morphologies were selected, and the same colony was streaked 3-5 more times to isolate and culture, ultimately obtaining two potential petroleum-degrading bacteria. The two obtained petroleum-degrading bacteria were then subjected to colony morphological characteristics and molecular biological identification. Identification showed that the obtained C1 colony was orange-red, opaque, and raised (e.g., ...). Figure 1(As shown); C3 colonies, color changes from white to green, opaque, raised (as shown). Figure 2 (As shown). A phylogenetic tree was constructed using MEGA 7.0 software based on the maximum likelihood method. After identification, they were named Gordonia sp. Strain C1 and Cellulosimicrobium sp. Strain C3, respectively.

[0050] The *Gordonia* sp. *Strain* C1 strain was deposited on September 15, 2022, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with accession number CGMCC No. 25708. The corresponding GenBank accession number is OP389054.1.

[0051] The fibrous microbacterium C3 (Cellulosimicrobium sp. Strain C3) was deposited on September 15, 2022, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, with accession number CGMCC No. 25709. The corresponding GenBank accession number is OP389052.1.

[0052] 1% (volume fraction for liquid culture medium and mass fraction for solid culture medium) of different carbon sources were added to the microbial enrichment medium to test the carbon source utilization of two strains. The detailed results are shown in Table 1, where “+” indicates strain growth and “-” indicates no growth.

[0053] Table 1 Carbon Source Utilization Table

[0054] carbon source Gordon's C1 Fiber microbes C3 glucose ﹣ ﹣ fructose ﹣ ﹣ sucrose ﹢ ﹢ Trehalose ﹢ ﹢ Cellobiose ﹣ ﹣ Sodium acetate ﹢ ﹢ Sodium pyruvate ﹢ ﹢ Sodium succinate ﹢ ﹢ Sodium fumarate ﹢ ﹣ Sodium citrate ﹣ ﹣ Sodium gluconate ﹢ ﹢ glycerin ﹢ ﹢ n-Butanol ﹢ ﹣ Twain 80 ﹢ ﹢ hexadecane ﹢ ﹢ paraffin oil ﹢ ﹢

[0055] Analysis of the carbon source utilization of different strains shows that *Gordonella* C1 and *Microsporum cytogenes* C3 have great application potential in alkane degradation, and also lay the foundation for designing specific nutrient culture formulations (such as modified BO medium). Furthermore, because the two strains utilize carbon sources differently, their combined application will have a wider range of carbon source utilization.

[0056] Example 2: Functional characterization of strains of Gordon's C1 and Microfibrillaria C3 cultured in biodegradable (BO) medium.

[0057] The functional traits of the strain mainly include growth potential, emulsifying properties, cell surface hydrophobic properties, and petroleum degradation potential.

[0058] The *Gordonia* sp. *Strain* C1 and *Cellulosimicrobium* sp. *Strain* C3 obtained in Example 1 were activated using LB1 medium and cultured at 37°C and 180 rpm for 60 hours. The cells were washed twice with 1×PBS, and the OD of the strain suspension was adjusted. 600 =1. Inoculate 1‰ of the bacterial culture into LB1 and BO media respectively, and incubate at 37℃ and 200 rpm for 144 hours. Take samples regularly to measure the OD of the bacterial culture. 600 The results showed that the strain grew relatively well in LB1 medium, but compared with BO medium, it took longer to reach the stationary phase and had a longer lag phase. Figure 3 In addition, the fibrous microbe C3 exhibits a relatively high growth rate.

[0059] On the other hand, the activated cell suspension was inoculated into BO medium at an inoculation rate of 1‰, and cultured at 26℃, 30℃, and 37℃, 200 rpm for 120 hours, respectively. Odion displacement (OD) of the bacterial suspension was measured periodically. 600 The results showed that the growth of the strain was relatively consistent at the three different temperatures. Figure 4 Furthermore, the growth of fibrous microbe C3 was relatively higher than that of Gordon's C1, indicating that the strain has the potential for room temperature application.

[0060] Through MATH experiment [1] The surface hydrophobicity of the cells was assessed, and the results are shown in Table 2. Table 2 shows that the surface hydrophobicity of *Gordonella* C1 was significantly higher than that of *Microbes fibrous* C3, especially under BO medium conditions, indicating that BO medium is most conducive to cell adhesion to petroleum hydrocarbon surfaces and degradation of petroleum hydrocarbons.

[0061] Table 2. Cell surface hydrophobicity (CSH%) of strains after culture in different culture media

[0062]

[0063] Surfactants enhance the solubility of petroleum hydrocarbon pollutants, promote the entry of hydrophobic substances into cells, and accelerate their biodegradation. Using paraffin oil as a crude oil substitute, the emulsifying performance of bacteria was assessed using the E24 value, reflecting the strain's ability to produce surfactants. Table 3 shows that the E24 value of *Microbes fibrocarpa* C3 is significantly higher than that of *Goldenella* C1, indicating its superior emulsifying performance.

[0064] Table 3. E24 content of supernatant after culturing in different culture media

[0065] LB1-E24 BO-E24 Gordon's C1 <![CDATA[12.66±2.51 b ]]> <![CDATA[6.67±2.89 b ]]> Fiber microbes C3 <![CDATA[75±5 a ]]> <![CDATA[34.33±4.04 a ]]>

[0066] The Gordonella C1 and Fibromicrobial C3 from Example 1 were inoculated at a rate of 15% into LB1 medium (petroleum content 80000 mg / L) and petroleum degradation medium, respectively, and cultured at 37°C and 200 rpm for 24 days.

[0067] Petroleum residues in the fermentation broth after biodegradation were extracted using ultrasonic extraction, and the total petroleum hydrocarbon degradation rate was determined by gravimetric method to evaluate the degradation performance of a single strain. [2] The result is as follows Figure 5 As shown, when using LB1 medium, the maximum degradation rate of total petroleum hydrocarbons was 43.36%, while when using petroleum degradation medium, the degradation rate of total petroleum hydrocarbons was greater than 60%.

[0068] Petroleum residues in the petroleum degradation medium were extracted with n-hexane and analyzed by GC-MS. [3],[4] The remaining C in the experimental group (group with added bacterial agent) and the control group (group without added bacterial agent) 10 -C 40 The aliphatic hydrocarbons were analyzed. The determination was performed according to the national standard "HJ1021-2019 Determination of Petroleum Hydrocarbons (C10-C40) in Soils and Sediments - Gas Chromatography". The results are as follows: Figure 6 As shown in the GC-MS peak diagram, the strain's response to C... 10 -C 40 All aliphatic hydrocarbons showed good degradation effects, C 10 -C 40 The degradation rates were 79.37% and 72.94%, respectively.

[0069] Example 2 demonstrates that the Gordonella C1 strain provided by this invention has a strong petroleum degradation capacity in biodegradable (BO) medium, and has good application prospects in petroleum microbial ecological restoration.

[0070] Examples 3-6: Evaluation of the petroleum degradation performance of two petroleum-degrading bacteria under different culture conditions

[0071] The implementation method is as described in Example 2. Different culture conditions were set, and the culture medium was inoculated with 15% of the culture medium. After 24 days of culture, the petroleum degradation performance of the single bacteria was evaluated by gravimetric method. The results are shown in Table 4.

[0072] Table 4 Total petroleum hydrocarbon degradation rate under different culture conditions

[0073]

[0074]

[0075] Under different culture conditions, the petroleum hydrocarbon degradation rate of both petroleum-degrading bacteria was over 60%, indicating that the single strain provided by this invention has flexible culture and application conditions.

[0076] Example 7: Evaluation of the petroleum degradation performance of the compound bacterial solution

[0077] The *Gordonia* sp. *Strain* C1 and *Cellulosimicrobium* sp. *Strain* C3 obtained in Example 1 were activated separately, washed, and the cell suspensions were adjusted to their OD values. 600 =1, mix Gordon's bacterium C1 and fibroblastic microbes C3 simultaneously at a volume ratio of 60% and 40% respectively to prepare a composite bacterial solution.

[0078] The compound bacterial culture was inoculated at a rate of 15% into a modified BO medium containing 80,000 mg / L petroleum. A control group was prepared using modified BO medium containing 80,000 mg / L petroleum but without the bacterial culture. The culture was carried out at 37℃ and 180-200 rpm for 24 days. Destructive sampling was performed on days 0, 8, 16, and 24. [3] .

[0079] The total petroleum hydrocarbon degradation rate of the compound bacterial solution on days 8, 16, and 24 was determined by gravimetric method. The results are as follows: Figure 7 As shown, the total petroleum hydrocarbon degradation rate was 48.43% on day 8, 69.36% on day 16, and reached 85.35% on day 24. These results indicate that the composite microbial community possesses a high petroleum degradation capacity and exhibits superior degradation performance compared to single strains. The cell surface hydrophobicity of *Gordonella* C1 is significantly higher than that of *Microbes fibrousa* C3, which facilitates cell adhesion to petroleum hydrocarbon surfaces and their degradation, demonstrating a strong petroleum pollution remediation capacity. Increasing its proportion further enhances the degradation effect. *Microbes fibrousa* C3 exhibits a relatively higher growth rate than *Gordonella* C1, promoting the degradation of some alkyl substances.

[0080] Examples 8-9: Effect of the proportion of single bacteria in the compound bacterial solution on the petroleum degradation rate

[0081] The implementation method is as described in Example 7. Different volume ratios of single bacteria in the compound bacterial solution were set. On the 8th, 16th and 24th days of cultivation, the ability of single bacteria in the compound bacterial solution with different volume ratios to degrade petroleum was evaluated by gravimetric method. The results are shown in Table 5.

[0082] Table 5 Total petroleum hydrocarbon degradation rate under different single-strain ratios.

[0083]

[0084] The varying volume percentages of individual bacteria in the composite microbial community had little impact on the final petroleum degradation performance. The total petroleum hydrocarbon degradation rate of the composite microbial community was consistently above 60%, with the highest reaching 86.52%, indicating that the composite microbial community provided by this invention offers flexible application options. Among the bacteria, the primary functional degrading bacterium was *Gordonella* C1, while the fibrous microbe C3 exhibited rapid growth, promoting degradation.

[0085] Examples 10-11: The ability of compound bacterial solution to treat different concentrations of petroleum

[0086] The implementation method is as described in Example 7. Different petroleum degradation concentrations were set, and the degradation performance of the compound bacterial solution on different petroleum concentrations was evaluated by gravimetric method on the 8th, 16th and 24th days of cultivation. The results are shown in Table 6.

[0087] Table 6 Total petroleum hydrocarbon degradation rate under different petroleum concentrations

[0088]

[0089] The total petroleum hydrocarbon degradation rate of the compound microbial community in modified BO medium (petroleum concentration of 60,000-100,000 mg / L) with added petroleum was 77.02%-86.15% after 24 days, indicating that the compound microbial community provided by the present invention has a high degradation rate, high concentration of petroleum pollution treatment, and a wide range of applications.

[0090] The specific embodiments described above further illustrate the purpose, technical solution, and beneficial effects of the present invention. It should be understood that the above descriptions are merely specific embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

[0091] References:

[0092] [1] Huang Xiangfeng, Fang Zheng, Huang Wei, Peng Kaiming, Lu Lijun, Liu Jia. Research progress on characterization of hydrophobicity of environmental microbial cell surface by MATH method [J]. Bulletin of Microbiology, 2015, 42(01):200-206;

[0093] [2] Bao Qinghua, Huang Lixin, Xiu Jianlong, Yi Lina. Advances in environmentally friendly treatment technology for oily sludge from oil and gas fields [J]. Applied Chemical Industry, 2021, 50(09):2602-2608;

[0094] [3]Geng Pengxue,Ma Anzhou,Wei Xiaoxia,Chen Xianke,Yin Jun,Hu Futang,Zhuang Xuliang,Song Maoyong,Zhuang Guoqiang. Interaction and spatio-taxonomic patterns of the soil microbiome around oil production wells impacted by petroleum hydrocarbons.[J].Environmental pollution(Barking,Essex:1987),2022,307;

[0095] [4] Guo Yan, Ma Jian, Yang Zongzheng, Zhang Tianyu, Sun Wei, Wu Zhiguo. Screening, identification and degradation characteristics of petroleum hydrocarbon degrading bacterium Mycolicibacterium fluoranthenivorans Y3 [J]. Environmental Science Guide, 2022, 41(05):1-7.

Claims

1. A bacterial community, characterized in that, The bacterial community comprises two organisms: Gordonia sp. C1 (accession number CGMCC No. 25708) and Cellulosimicrobium sp. C3 (accession number CGMCC No. 25709). The bacterial community consists of Gordonia sp. C1 comprising 50%-70% of the volume and Cellulosimicrobium sp. C3 comprising 30%-50% of the volume.

2. The method for culturing the microbial community according to claim 1, characterized in that, The culture medium used for cultivation consisted of the following components: CH3COONa 1 g / L, C3H3NaO3 0.5 g / L, K2HPO4 0.5 g / L, KNO3 0.1 g / L, NH4Cl 0.05 g / L, NaCl 25 g / L, FeC6H5O7 0.002 g / L, and natural pH. The cultivation conditions were: 26℃-37℃, 150-200 rpm.

3. The application of the microbial community described in claim 1 in the field of oil pollution environmental remediation.

4. The application according to claim 3, wherein the oil-polluted environment includes oil-polluted soil and oil-polluted water.

5. A method for microecological remediation of petroleum pollution, characterized in that, The method described in claim 1 uses the microbial community to degrade petroleum hydrocarbons in petroleum-contaminated soil or water.

6. A method for degrading petroleum hydrocarbons, characterized in that, The method involves inoculating the bacterial community of claim 1 into a degradation system containing petroleum at a mass concentration of 60,000-100,000 mg / L and culturing it at 26℃-37℃ and 150-200 rpm.

7. The method according to claim 6, characterized in that, The nutrients in the degradation system are CH3COONa 0.3 g / L, C3H3NaO3 0.1 g / L, K2HPO4 1 g / L, KNO3 2 g / L, NH4Cl 0.5 g / L, NaCl 10 g / L, and FeC6H5O7 0.005 g / L.

8. An article for treating petroleum-polluted environments, characterized in that, The product contains the microbial community as described in claim 1.

9. The article of claim 8, characterized in that, The product is a liquid microbial agent.

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