Rhodococcus capable of stably and efficiently degrading estrogen and application of rhodococcus

Abstract

The invention relates to rhodococcus capable of stably and efficiently degrading estrogen and application of the rhodococcus. The rhodococcus C1 is preserved in the China General Microbiological Culture Collection Center on May 15, 2023, the preservation address is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the preservation number is CGMCC No.27337. Compared with the prior art, the Rhodococcus sp C1 provided by the invention can efficiently degrade the natural estrogen 17-beta estradiol, is tolerant to high-salt and heavy metal ions, can stably degrade the 17-beta estradiol, and has the application potential of repairing estrogen pollution.

Description

Technical Field

[0001] The present invention relates to the technical field of microorganisms, and in particular to a Rhodococcus bacterium capable of stably and efficiently degrading estrogen and its application. Background Art

[0002] Environmental estrogens are a new type of pollutant that harms the normal hormone functions and endocrine systems of humans, animals and plants. They have carcinogenic, teratogenic and mutagenic effects on organisms. Trace estrogen compounds can affect the metabolism of organisms. By interfering with and antagonizing the normal synthesis, transportation and release of endogenous hormones in organisms, they can cause disorders in the hormone balance and regulation systems of humans and animals, resulting in feminization of male organisms, lesions of reproductive organs, and an increase in the incidence of cancer. Therefore, estrogen compounds have been listed as a class of carcinogens by the World Health Organization. Environmental estrogen pollutants are difficult to decompose, remain in the environment for a long time, accumulate and amplify, and are widely distributed in various habitats such as human and animal excreta, waste from farms, wastewater from sewage treatment plants, and farmland soil. Environmental estrogen pollution has become a global environmental problem.

[0003] Natural estrogens are the main components of environmental estrogen pollutants, mainly including estrone (E1), 17β-estradiol (E2) and estriol (E3) compounds. Among them, 17β-estradiol (E2) is currently the natural estrogen with the highest activity and the strongest endocrine disrupting effect, and its activity intensity is 5-1000 times that of estrone and estriol. At present, the traditional physical and chemical methods for treating environmental estrogen pollutants have unsatisfactory effects, high costs, slow effects, many secondary products, and poor sustainability. Biodegradation methods, especially microbial degradation methods, have significant advantages such as high efficiency, low energy consumption, simple operation, and no secondary pollution, and have become the focus and key point in the research of environmental pollutant removal and environmental remediation.

[0004] Microorganisms have strong environmental adaptability and high metabolic diversity, and can degrade a variety of inorganic and organic substances, with great application potential. However, the degradation efficiency of most degradable microorganisms is unstable and their environmental tolerance is poor, which greatly limits their practical applications. Many estrogen-degrading strains isolated in laboratories often have poor environmental adaptability. Many factors in the actual environment, such as pH, salinity, other carbon sources, and co-existing other pollutants, such as surfactants and heavy metals, will affect the survival and degradation of microorganisms. At present, the concentration ranges of heavy metal ions Cu 2+ and Cd 2+ in the actual livestock and poultry waste environment in China are concentrated in the ranges of 1.92-5.78 mg / L and 0.02-2.72 mg / L respectively, and the salt concentration is mainly concentrated in the range of 1.8-24.2 g / kg. Therefore, microbial strains with good environmental adaptability and capable of stably and efficiently degrading pollutant estrogens will promote the efficient removal of estrogen pollutants and environmental bioremediation, and promote the stability and sustainable development of the ecological environment.

[0005] Chinese Patent CN104894012A discloses a Rhodococcus strain capable of degrading 17β-estradiol, named JX-2, with a deposit number of CGMCC NO.9636. This solution uses Rhodococcus JX-2 to produce an immobilized microbial agent for degrading 17-β estradiol. By uniformly mixing Rhodococcus JX-2 with sodium alginate solution at a volume ratio of 1 - 1.5:1 and using a 5% mass concentration of CaCl2 solution as a cross-linking agent, immobilized beads with a diameter of 3 - 5 mm are obtained after cross-linking for 5.5 - 6.5 hours. Rhodococcus JX-2 can grow with E2 as the sole carbon source under shake flask culture conditions and degrade more than 90% of 30 mg / L of 17-β estradiol within 7 days. The immobilized microbial agent using this strain can degrade more than 60% of 17-β estradiol in sewage within 7 days and more than 80% of 17-β estradiol in livestock manure within 7 days.

[0006] Chinese Patent CN112410245A discloses a low-temperature composting microbial composite agent and its preparation method and application. The invention relates to a low-temperature composting microbial composite agent for promoting the degradation of agricultural waste in cold regions, including a cold-tolerant cellulose-degrading microbial agent composed of two cold-tolerant cellulose-degrading bacteria, and a steroid estrogen-degrading microbial agent composed of a 17β-estradiol-degrading Rhodococcus and a estrone-degrading Microbacterium. The 17β-estradiol-degrading bacterium described in this solution uses E2 as the sole carbon source, and the highest degradation rate of 30 mg / L E2 is 97.98% on the 3.5th day.

[0007] Chinese Patent CN101705195A discloses a Bacillus strain capable of degrading 17β-estradiol, named E2-Y, with a strain deposit number of CCTCC M209150. The strain E2-Y of this invention can completely remove ≤10 mg / L of 17-β estradiol within 5 days and completely remove ≤50 mg / L of 17-β estradiol within 7 days. The strain E2-Y has good acid-base tolerance. When the pH is 6.5 - 8.5, it can degrade more than 65% of 17-β estradiol, and the degradation rate of 17-β estradiol can reach 90% when the pH is 7.5.

[0008] A Rhodococcus strain that grows with E2 as the sole carbon source and energy source was screened from Genome Analysis of Rhodococcus Sp.DSSKP-R-001:A Highly Effectiveβ-Estradiol-Degrading Bacterium (Zhao etal.2018) and named DSSKP-R-001. The degradation rate of this strain for 50 mg / L E2 was 97% on the 3rd day.

[0009] Since most of the actual polluted environments where estrogen pollutants are located are characterized by high salinity, high heavy metals, and mixed pollutants, the isolated estrogen-degrading strains should not only have stable degradation efficiency but also possess good environmental tolerance to achieve effective microbial remediation of the actual complex polluted environment. However, the Rhodococcus discovered in the above prior art has not explored its tolerance to actual complex polluted environments such as high salinity and heavy metals, and the estrogen pollution concentration it can degrade is low, the stress conditions it can tolerate are few, the degradation efficiency is low, and the repair cycle is long. Summary of the Invention

[0010] The object of the present invention is to provide a Rhodococcus and its application for stably and efficiently degrading estrogen. The Rhodococcus (Rhodococcus.sp) C1 provided by the present invention can efficiently degrade natural estrogen 17-β estradiol, tolerate high salinity and heavy metal ions, and can stably degrade 17-β estradiol, having the application potential for repairing estrogen pollution.

[0011] The object of the present invention can be achieved by the following technical solutions:

[0012] A Rhodococcus C1 for stably and efficiently degrading estrogen was deposited with the China Center for Type Culture Collection on May 15, 2023. The deposit address is No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC NO. 27337.

[0013] Furthermore, the 16S rDNA sequence of the Rhodococcus C1 is as shown in SEQ ID NO.1.

[0014] The Rhodococcus C1 provided by the present invention was obtained by multiple rounds of enrichment and screening from polluted soil. The colony of strain C1 is round, raised, light yellow, opaque, Gram-positive, aerobic, and grows well at 30°C; 16S rDNA gene sequence analysis shows that this strain C1 is Rhodococcus (Rhodococcus.sp).

[0015] The present invention also provides a microbial preparation containing the above Rhodococcus C1.

[0016] Furthermore, in the microbial preparation, the concentration of Rhodococcus C1 is at least 10 OD 600 / L.

[0017] Furthermore, the microbial preparation is a solid, liquid bacterial agent or freeze-dried powder.

[0018] Furthermore, the microbial preparation also includes ethanol, glucose, glycerol, etc., which assist in degrading estrogen through composite reagents and can improve the degradation efficiency.

[0019] As a preferred technical solution, the mass percentage of the ethanol is 0.1% (w / w%).

[0020] The present invention also provides a product, which contains the above-mentioned Rhodococcus sp. C1 or contains the above-mentioned microbial preparation.

[0021] Furthermore, in the product, the concentration of Rhodococcus sp. C1 is at least 10 OD 600 / L.

[0022] Furthermore, the product is a feed additive or a chemical.

[0023] The present invention also provides an application of the above-mentioned Rhodococcus sp. C1, or the above-mentioned microbial preparation, or the above-mentioned product in degrading estrogen.

[0024] Furthermore, the estrogen is 17β-estradiol (E2).

[0025] The present invention also provides an application of the above-mentioned Rhodococcus sp. C1, or the above-mentioned microbial preparation, or the above-mentioned product in degrading 17β-estradiol under high-salt and heavy-metal conditions.

[0026] Furthermore, in the conditions, the NaCl concentration range is 10-40 g / L; Cu 2+ concentration range is 1.25-6.25 mg / L, Cd 2+ concentration range is 1.25-6.25 mg / L.

[0027] Furthermore, the high-salt and heavy-metal environment includes water environment, soil environment, etc.

[0028] Furthermore, in the water environment, the concentration of 17β-estradiol is 10 mg / L to 50 mg / L.

[0029] As a preferred technical solution, the Rhodococcus sp. C1 of the present invention can efficiently degrade 17β-estradiol, and the degradation rates of 17β-estradiol in water bodies with 17β-estradiol concentrations of 10 mg / L, 20 mg / L, and 50 mg / L within 48 hours are 98.6%, 98.1%, and 93.9% respectively.

[0030] Furthermore, in the soil environment, the concentration of 17β-estradiol is 10 mg / kg to 50 mg / kg.

[0031] As a preferred technical solution, the Rhodococcus sp. C1 of the present invention can efficiently degrade 17β-estradiol, and the degradation rate of 17β-estradiol in soil with a 17β-estradiol concentration of 10 mg / kg within 24 hours is 92%.

[0032] In addition, the present invention also provides a method for obtaining Rhodococcus sp. C1 that can stably and efficiently degrade estrogen, and the specific steps are as follows:

[0033] S1. Collect contaminated soil as a microbial source;

[0034] S2. Prepare an inorganic salt medium containing 10 mg / L of 17β-estradiol (E2), adjust the pH value to 7.4 with hydrochloric acid, and sterilize at 121 °C for 20 min;

[0035] S3. When the temperature of the medium drops to 30 °C ± 1 °C, inoculate the soil under the laminar flow hood at an inoculation amount of 10%;

[0036] S4. Incubate in a constant temperature shaker incubator at 30 °C and 180 r / min for enrichment culture for 7 days;

[0037] S5. After repeating the enrichment culture 5 times, streak culture repeatedly on a solid medium plate until a purified monoclonal strain is isolated and named Rhodococcus sp. C1.

[0038] Further, in step S2, the components and concentrations of the inorganic salt medium are as follows: K2HPO4·3H2O, 13.75 g / L; KH2PO4, 4.5 g / L; (NH4)2SO4, 2.0 g / L; MgSO4·7H2O, 0.16 g / L; FeSO4·7H2O, 0.005 g / L; CaCl 2· 2H2O, 0.011 g / L; MnCl2·4H2O, 0.002 g / L.

[0039] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0040] The Rhodococcus sp. C1 provided by the present invention can efficiently degrade the natural estrogen 17β-estradiol, has good adaptability to complex environments such as high salt and heavy metals, and can be applied to the effective removal and bioremediation of natural estrogen pollutants in environments such as water bodies and soils.

[0041] The Rhodococcus sp. C1 provided by the present invention can use the natural estrogen 17β-estradiol (E2) as a carbon source, and the degradation rates of 10 mg / L, 20 mg / L, and 50 mg / L of 17β-estradiol within 48 hours are 98.6%, 98.1%, and 93.9% respectively. In simulated soil, the degradation rate of Rhodococcus sp. C1 to 10 mg / kg of 17β-estradiol within 24 hours is 92%. Strain C1 can tolerate 40 g / L of NaCl, 6.25 mg / L of Cu 2+ and 6.25 mg / L of Cd 2+ , and can stably degrade 17β-estradiol.

[0042] Compared with other discovered natural estrogen-degrading strains, the strain of the present invention, Rhodococcus sp. C1, has a stronger tolerance concentration to 17-β estradiol, a higher degradation efficiency, a shorter degradation period, more stable degradation in water and soil, a stronger environmental adaptability, and a wider range of actual environments to which it can be applied. Therefore, Rhodococcus sp. C1 provided by the present invention has better efficacy and potential in terms of natural estrogen degradation efficiency, degradation period, and estrogen repair effect in the actual complex polluted environment. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 It is a spread plate diagram of LB plate of Rhodococcus sp. C1 of the present invention;

[0044] Figure 2 It is a schematic diagram of the degradation curve of Rhodococcus sp. C1 of the present invention for 17β-estradiol with different concentrations (10mg / L, 20mg / L, and 50mg / L) in water;

[0045] Figure 3 It is a schematic diagram of the growth of Rhodococcus sp. C1 of the present invention in water containing different concentrations (10mg / L, 20mg / L, and 50mg / L) of 17β-estradiol;

[0046] Figure 4 It is a schematic diagram of the degradation curve of Rhodococcus sp. C1 of the present invention for 17β-estradiol in soil containing 10mg / kg of 17β-estradiol;

[0047] Figure 5 It is for Rhodococcus sp. C1 of the present invention for different concentrations (0mg / L, 1.25mg / L, 6.25mg / L) of Cu 2+ and the degradation curve schematic diagram of 17β-estradiol in the inorganic salt medium with 10mg / L of 17β-estradiol;

[0048] Figure 6 It is for Rhodococcus sp. C1 of the present invention in different concentrations (0mg / L, 1.25mg / L, 6.25mg / L) of Cu 2+ and the growth situation schematic diagram in the inorganic salt medium with 10mg / L of 17β-estradiol;

[0049] Figure 7 It is a schematic diagram of the degradation curve of Rhodococcus sp. C1 of the present invention for 17β-estradiol in the inorganic salt medium with different concentrations (0g / L, 20g / L, 40g / L, 80g / L) of sodium chloride and 10mg / L of 17β-estradiol;

[0050] Figure 8 Schematic diagram of the growth of Rhodococcus sp. C1 of the present invention in an inorganic salt medium with different concentrations (0 g / L, 20 g / L, 40 g / L, 80 g / L) of sodium chloride and 10 mg / L of 17β-estradiol;

[0051] Figure 9 Schematic diagram of the growth of Rhodococcus sp. C1 of the present invention in an LB medium with different concentrations (10 g / L, 20 g / L, 40 g / L, 80 g / L) of sodium chloride and 10 mg / L of 17β-estradiol;

[0052] Figure 10 Schematic diagram of the growth of Rhodococcus sp. C1 of the present invention in an LB medium with different concentrations (0 mg / L, 0.05 mg / L, 0.25 mg / L, 1.25 mg / L, 6.25 mg / L, 31.25 mg / L) of Cd 2+ and 10 mg / L of 17β-estradiol. Detailed implementation manners

[0053] The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments. This embodiment is implemented on the premise of the technical solution of the present invention, and the detailed implementation manners and specific operation processes are given, but the protection scope of the present invention is not limited to the following embodiments.

[0054] In the following embodiments, the 16S rDNA sequence of Rhodococcus C1 is as shown in SEQ ID NO.1:

[0055]

[0056] The following will, in conjunction with the accompanying drawings, elaborate on some embodiments of the present invention. Without conflict, the following embodiments and the features in the embodiments can be combined with each other.

[0057] Example 1

[0058] This example provides a method for obtaining Rhodococcus sp. C1, and the specific steps are as follows:

[0059] S1. Prepare an inorganic salt medium containing 10 mg / L of 17β-estradiol (E2). The components of the inorganic salt medium are as follows: K2HPO4·3H2O, 13.75 g / L; KH2PO4, 4.5 g / L; (NH4)2SO4, 2.0 g / L; MgSO4·7H2O, 0.16 g / L; FeSO4·7H2O, 0.005 g / L; CaCl 2· 2H2O, 0.011 g / L; MnCl2·4H2O, 0.002 g / L. Adjust the pH value to 7.4 with hydrochloric acid, and sterilize it at 121 °C for 20 min in an autoclave for later use;

[0060] S2. Collect a petroleum-contaminated soil sample, and inoculate the soil sample into a 250 mL Erlenmeyer flask containing 100 mL of the inorganic salt medium prepared in step S1 at an inoculation amount of 10%, and enrich and culture it in a constant temperature shaker incubator at 30 °C and 180 r / min for 7 days;

[0061] S3. After repeating the enrichment culture 5 times, streak culture on an LB solid medium plate until purified single colonies are isolated and named C1.

[0062] As Figure 1 shown, the colonies of strain C1 are round, raised, light yellow, opaque, Gram-positive, aerobic, and grow well at 30 °C.

[0063] Example 2

[0064] This example provides a method for identifying Rhodococcus sp. C1. The 16S rDNA gene sequence of strain C1 screened in Example 1 is amplified, determined, and compared with the database. The specific steps are as follows:

[0065] S1. Use a genomic DNA extraction kit (FastPure Bacteria DNA Isolation Mini Kit - BOX2) to extract the genomic DNA of strain C1;

[0066] S2. Use a 16S rDNA fragment amplification and detection kit (MicroSEQTM Using the whole-genome 16S rDNA PCR kit, the genomic DNA of strain C1 was used as a template to amplify the 16S rDNA fragment of strain C1, and its gene sequence was determined.

[0067] The 16S rDNA sequence of strain C1 is shown in SEQ ID NO.1. Alignment with the NCBI database (http: / / www.ncbi.nlm.nih.gov / ) indicated that strain C1 belongs to Rhodococcus sp. The Rhodococcus C1 was deposited at the China General Microbiological Culture Collection Center on May 15, 2023. The deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC NO.27337.

[0068] Example 3

[0069] This example provides a test experiment on the degradation ability of Rhodococcus C1 to 17β-estradiol at different concentrations. The specific steps are as follows:

[0070] S1. Streak the Rhodococcus sp. C1 screened in Example 1 on an LB plate and culture it at 30°C for 2 days.

[0071] S2. Pick a single colony from the streaked plate obtained in step S1 and inoculate it into 3 ml of LB medium, and culture it overnight in a shaker at 30°C.

[0072] S3. Transfer the overnight-cultured bacterial liquid to a liquid LB medium at a ratio of 1:100 and continue to culture it until the logarithmic phase.

[0073] S4. Centrifuge the bacterial liquid in the logarithmic phase obtained in step S3 at 8000 rpm for 5 minutes at room temperature to collect the bacterial cell precipitate.

[0074] S5. Resuspend the bacterial cells using an inorganic salt medium (K2HPO4·3H2O, 13.75 g / L; KH2PO4, 4.5 g / L; (NH4)2SO4, 2.0 g / L; MgSO4·7H2O, 0.16 g / L; FeSO4·7H2O, 0.005 g / L; CaCl 2· 2H2O, 0.011 g / L; MnCl2·4H2O, 0.002 g / L, pH 7.4), and centrifuge and wash three times to remove the residual medium.

[0075] S6. Resuspend the bacterial cells in the inorganic salt medium, adjust OD 600 = 0.5, add 17β-estradiol at different concentrations (10 mg / L, 20 mg / L, 50 mg / L) to the medium, and culture it in a shaker at 30°C for 2 days.

[0076] S7. Add an equal volume of ethyl acetate to the culture, mix well and extract three times. After rotary evaporation to remove ethyl acetate, redissolve with acetonitrile.

[0077] S8. Take 10 μL of the extract and use high performance liquid chromatography to determine the residual amount of 17β-estradiol. The detection parameters are as follows: Agilent ZORBAX StableBond SB-C18 (4.6×150 mM, 5 μm) reversed-phase liquid chromatography column. The mobile phase volume ratio is acetonitrile: water (50:50, v / v), the flow rate is 1 mL / min, and the detection temperature is 30 °C. Use an FLD detector, and the excitation wavelength / emission wavelength is 280 / 315 nm.

[0078] The results are as Figures 2 - 3 shown. Strain C1 can use 17β-estradiol as a carbon source, and the degradation rates of 17β-estradiol at 10 mg / L, 20 mg / L and 50 mg / L within 48 hours are 98.6%, 98.1% and 93.9% respectively. Therefore, Rhodococcus sp. C1 can efficiently degrade 17β-estradiol in water bodies.

[0079] Example 4

[0080] This example provides a test experiment on the degradation ability of Rhodococcus sp. C1 to 17β-estradiol in soil. The specific steps are as follows:

[0081] S1. Streak the Rhodococcus sp. C1 screened in Example 1 on an LB plate and culture at 30 °C for 2 days.

[0082] S2. Pick monoclonal colonies from the streaked plate obtained in step S1 and inoculate them into 3 ml of LB medium, and culture overnight in a shaker at 30 °C.

[0083] S3. Transfer the overnight culture broth to a liquid LB medium at a ratio of 1:100 and continue to culture until the logarithmic phase.

[0084] S4. Centrifuge the broth in the logarithmic phase obtained in step S3 at 8000 rpm for 5 minutes at room temperature to collect the cell pellet.

[0085] S5. Resuspend the cells using an inorganic salt medium (K2HPO4·3H2O, 13.75 g / L; KH2PO4, 4.5 g / L; (NH4)2SO4, 2.0 g / L; MgSO4·7H2O, 0.16 g / L; FeSO4·7H2O, 0.005 g / L; CaCl 2· 2H2O, 0.011 g / L; MnCl2·4H2O, 0.002 g / L, pH 7.4), and centrifuge and wash three times to remove the residual medium.

[0086] S6. Resuspend the bacterial cells in an inorganic salt medium and adjust the OD 600 value to 2.0. Inoculate the bacterial solution into 200 g of soil at a ratio of 5 OD 600 / 100 g of soil, add 17β-estradiol to a final concentration of 10 mg / kg, and culture at 30 °C for 10 days. The soil inoculated with the heat-sterilized strain C1 is set as a blank control to evaluate its abiotic loss.

[0087] S7. Use ethyl acetate to extract 17β-estradiol from the soil sample by shaking, repeat three times, remove ethyl acetate by rotary evaporation, and then redissolve with acetonitrile.

[0088] S8. Take 10 μL of the extract and determine the residual amount of 17β-estradiol by high performance liquid chromatography.

[0089] The results are as Figure 4 shown. In the simulated soil environment, the degradation rate of strain C1 to 10 mg / kg of 17β-estradiol within 24 hours is 92%. Therefore, Rhodococcus sp. C1 can efficiently degrade 17β-estradiol in soil.

[0090] Example 5

[0091] This example provides an environmental adaptability detection experiment of Rhodococcus sp. C1, and the specific steps are as follows:

[0092] S1. Streak the Rhodococcus sp. C1 screened in Example 1 on an LB plate and culture at 30 °C for 2 days.

[0093] S2. Pick a single colony from the streaked plate obtained in step S1 and inoculate it into 3 ml of LB medium, and culture overnight in a shaker at 30 °C.

[0094] S3. Collect the culture, wash it, and then resuspend and inoculate it into an inorganic salt medium (K2HPO4·3H2O, 13.75 g / L; KH2PO4, 4.5 g / L; (NH4)2SO4, 2.0 g / L; MgSO4·7H2O, 0.16 g / L; FeSO4·7H2O, 0.005 g / L; CaCl 2+ containing 10 mg / L of 17β-estradiol and different concentrations (10 g / L, 20 g / L, 40 g / L, 60 g / L) of sodium chloride, different concentrations (0 mg / L, 1.25 mg / L, 6.25 mg / L) of Cu 2+ or different concentrations (0 mg / L, 0.05 mg / L, 0.25 mg / L, 1.25 mg / L, 6.25 mg / L, 31.25 mg / L) of Cd 2·2H2O, 0.011 g / L; MnCl2·4H2O, 0.002 g / L, pH 7.4), adjust the bacterial amount to 0.05 OD 600 / mL, culture in a shaker at 30 °C for 2 days, and detect OD 600 and the residual amount of 17β-estradiol.

[0095] The results are as Figures 5 - 10 shown. Strain C1 can grow normally in the medium containing 40 g / L NaCl, 6.25 mg / L Cu 2+ and 6.25 mg / L Cd 2+ and can still stably and effectively degrade 17β-estradiol in the presence of 40 g / L NaCl, 6.25 mg / L Cu 2+ .

[0096] The above description of the embodiments is to enable those of ordinary skill in the art to understand and use the invention. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative efforts. Therefore, the present invention is not limited to the above embodiments, and all improvements and modifications made by those skilled in the art without departing from the scope of the present invention should be within the protection scope of the present invention.

Claims

1. Rhodococcus sp. C1 for stably and efficiently degrading estrogen, characterized in that The Rhodococcus sp. C1 was deposited at the China General Microbiological Culture Collection Center on May 15, 2023. The deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC NO. 27337.

2. The Rhodococcus sp. C1 for stably and efficiently degrading estrogen according to claim 1, characterized in that The 16S rDNA sequence of the Rhodococcus sp. C1 is shown as SEQ ID NO.

1.

3. A microbial preparation containing the Rhodococcus sp. C1 as claimed in claim 1 or claim 2.

4. The microbial agent according to claim 3, characterized in that, In the microbial agent, the concentration of Rhodococcus sp. C1 is at least 10 OD 600 / L.

5. A product, characterized in that, The product contains the Rhodococcus sp. C1 as claimed in claim 1 or claim 2, or contains the microbial preparation as claimed in claim 3 or claim 4.

6. The product according to claim 5, wherein, In the said product, the concentration of Rhodococcus sp. C1 is at least 10 OD 600 / L.

7. Use of the Rhodococcus sp. C1 as claimed in claim 1 or claim 2, or the microbial preparation as claimed in claim 3 or claim 4, or the product as claimed in claim 5 or claim 6 in degrading estrogen.

8. The application according to claim 7, characterized in that The estrogen is 17β-estradiol.

9. Use of the Rhodococcus sp. C1 as claimed in claim 1 or claim 2, or the microbial preparation as claimed in claim 3 or claim 4, or the product as claimed in claim 5 or claim 6 in degrading 17β-estradiol under high salt and heavy metal conditions.

10. The application according to claim 9, wherein, The NaCl concentration range in the said conditions is 10 - 40 g / L; Cu 2+ concentration range is 1.25 - 6.25 mg / L, Cd 2+ concentration range is 1.25 - 6.25 mg / L.

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