Rosetonia for degrading pyridine

Abstract

The invention relates to Rosetonia for degrading pyridine, belongs to the technical field of microorganisms, and is used for solving at least one of the problems that the existing conventional microorganisms are difficult to degrade pyridine, the degradation efficiency is low and the like. The Rosetonia Q6 disclosed by the invention can realize efficient growth and degradation in an inorganic salt culture medium taking pyridine as a unique carbon source and energy source. The Ralstonia Q6 strain shows remarkable tolerance and decomposition capability on high-concentration pyridine pollutants, can effectively decompose pyridine pollutants in actual wastewater, and has a relatively good engineering application value in the field of treatment of high-concentration pyridine wastewater.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, and more particularly to a Rolstonia bacterium for degrading pyridine. Background Technology

[0002] Pyridine, a typical nitrogen-containing heterocyclic aromatic hydrocarbon, is widely found in the wastewater of numerous industries, including coal chemical industry, petroleum refining, pesticide synthesis, and pharmaceutical manufacturing. The nitrogen atom in its molecular structure endows pyridine with significant water solubility, making it highly susceptible to migration and diffusion with industrial wastewater, polluting surface water bodies and groundwater systems. Pyridine has been proven to possess high biotoxicity, potential carcinogenicity, and mutagenicity, posing a direct threat to aquatic organisms and posing a significant threat to human health and safety through bioaccumulation in the food chain or via drinking water. Furthermore, the persistence of pyridine in the environment exacerbates its ecological risks, making the effective control and elimination of pyridine pollution an urgent environmental problem to be solved.

[0003] Biological treatment technology is considered an ideal choice for treating wastewater containing organic pollutants such as pyridine due to its environmental friendliness, relatively low operating costs, and low risk of secondary pollution. This technology primarily relies on specific microbial populations, utilizing their metabolic activity to ultimately convert complex organic pollutants into harmless or low-toxicity inorganic small molecules (such as CO2, H2O, and NH4). + Biodegradation processes typically do not produce hazardous chemical byproducts requiring subsequent treatment or large amounts of chemical sludge, compared to chemical methods such as physical adsorption or advanced oxidation. This aligns with the principles of sustainable development and green governance.

[0004] However, the stability of pyridine's structure makes it a stubborn pollutant that is difficult to overcome in biodegradation systems. Conventional environmental microbial communities or existing engineered strains often have limited efficiency in degrading pyridine, failing to meet the stringent requirements for rapid and thorough purification of high-concentration pyridine wastewater. This insufficient degradation efficiency restricts the widespread application of biological treatment technologies in the field of pyridine pollution control. Summary of the Invention

[0005] In view of the above analysis, the present invention aims to provide a Roldstonia bacterium for degrading pyridine, in order to solve at least one of the problems of existing conventional microorganisms being unable to degrade pyridine and having low degradation efficiency.

[0006] In a first aspect, the present invention provides a Ralstonia sp. Q6 with accession number CGMCC No.33402.

[0007] Furthermore, the Rolstonia Q6 contains the 16S rDNA shown in SEQ ID No. 1.

[0008] Furthermore, the Rolstonia Q6 was isolated from oilfield sludge in an oil production plant.

[0009] Furthermore, the Ralstonia Q6 strain achieved a 100% removal rate of pyridine at concentrations ≤400 mg / L within a 72-hour treatment cycle, and removal rates of ≥98.3%, ≥86.2%, and ≥73.5% for pyridine at concentrations of 600 mg / L, 800 mg / L, and 1000 mg / L, respectively.

[0010] Secondly, the present invention provides a formulation for degrading pyridine, comprising the aforementioned Rolstonia Q6.

[0011] Thirdly, the present invention provides a method for degrading pyridine using the aforementioned Roldstone bacteria Q6, comprising the following steps:

[0012] (1) Expand the culture of Rolstonia Q6, centrifuge to obtain a bacterial suspension;

[0013] (2) Dissolve pyridine in an inorganic salt culture medium to obtain a culture medium containing pyridine;

[0014] (3) The bacterial suspension is added to the culture medium containing pyridine and cultured to degrade the pyridine.

[0015] Further, in step (1), the expanded culture specifically includes: inoculating the Ralstonia Q6 into LB medium, culturing it at 35-37°C and 120-150 r / min to the logarithmic growth phase, then collecting the bacterial cells by centrifugation at 5000-8000 r / min for 5-10 min, washing the bacterial cells with sterile physiological saline and resuspending them to an appropriate concentration to obtain the bacterial suspension.

[0016] Furthermore, in step (2), the concentration of pyridine in the pyridine-containing culture medium is ≥200 mg / L.

[0017] Furthermore, in step (3), the culture is carried out under the conditions of 35-37℃ and 120-180r / min in the dark with shaking, and the culture time is 72-80h.

[0018] Fourthly, the present invention provides an application of the above-mentioned Rolstonia Q6 in the degradation of pyridine-containing wastewater.

[0019] Compared with the prior art, the present invention can achieve at least one of the following beneficial effects:

[0020] 1. The Ralstonia Q6 strain of the present invention can achieve efficient growth and degradation in an inorganic salt culture medium with pyridine as the sole carbon and energy source. The Ralstonia Q6 strain exhibits significant tolerance and decomposition ability to high concentrations of pyridine pollutants, and can effectively decompose pyridine pollutants in actual wastewater, demonstrating good engineering application value in the treatment of high-concentration pyridine wastewater.

[0021] 2. The Ralstonia Q6 strain described in this invention achieves a 100% removal rate of pyridine at concentrations ≤400 mg / L within a 72-hour treatment cycle, and removal rates of ≥98.3%, ≥86.2%, and ≥73.5% for pyridine at concentrations of 600 mg / L, 800 mg / L, and 1000 mg / L, respectively.

[0022] 3. The Rolstonia Q6 strain of the present invention is suitable for the bioremediation of industrial wastewater, groundwater and contaminated soil containing pyridine. It exhibits significant tolerance and decomposition ability to high concentrations of pyridine pollutants and can effectively decompose pyridine pollutants in actual wastewater. It has good engineering application value in the field of treating high concentration pyridine wastewater.

[0023] In this invention, the above-described technical solutions can be combined with each other to achieve more preferred combinations. Other features and advantages of this invention will be set forth in the following description, and some advantages may become apparent from the description or be learned by practicing the invention. The objects and other advantages of this invention can be realized and obtained from what is particularly pointed out in the description and drawings. Attached Figure Description

[0024] The accompanying drawings are for illustrative purposes only and are not intended to limit the invention. Throughout the drawings, the same reference numerals denote the same parts.

[0025] Figure 1 The colony morphology characteristics of the Q6 strain of the present invention in inorganic salt solid culture medium;

[0026] Figure 2 Gram staining diagram of the Q6 strain of the present invention;

[0027] Figure 3 This is a scanning electron microscope image of the Q6 strain of the present invention;

[0028] Figure 4 This is the phylogenetic tree of the Q6 strain of the present invention;

[0029] Figure 5 This is a graph showing the pyridine removal rate of a Q6 strain of the present invention at different concentrations;

[0030] Figure 6 The degradation curve of the Q6 strain of the present invention using 600 mg / L pyridine as the sole carbon source is shown.

[0031] Figure 7 The cell growth curve of the Q6 strain of the present invention using 600 mg / L pyridine as the sole carbon source is shown.

[0032] Figure 8 The removal rate of pyridine by the Q6 strain of the present invention at different NaCl concentrations;

[0033] Figure 9 The figure shows the naphthalene removal rate of the Q6 strain of the present invention at different NaCl concentrations. Detailed Implementation

[0034] The preferred embodiments of the present invention will now be described in detail with reference to the accompanying drawings, which constitute a part of the present invention and are used together with the embodiments of the present invention to illustrate the principles of the present invention, but are not intended to limit the scope of the present invention.

[0035] In a specific embodiment of the present invention, a strain of Ralstonia sp. Q6 with accession number CGMCC No. 33402 is deposited at the China General Microbiological Culture Collection Center (CGMCC, address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences), with accession number CGMCC No. 33402 and accession date of January 14, 2025.

[0036] Compared with existing technologies, the Ralstonia Q6 strain of this invention can achieve efficient growth and degradation in inorganic salt culture media with pyridine as the sole carbon and energy source. The Ralstonia Q6 strain exhibits significant tolerance and decomposition ability to high concentrations of pyridine pollutants, effectively decomposing pyridine pollutants in actual wastewater, and has good engineering application value in the treatment of high-concentration pyridine wastewater.

[0037] Specifically, the Rolstonia Q6 contains the 16S rDNA shown in SEQ ID No. 1.

[0038] Specifically, the Rolstonia Q6 was isolated from oilfield sludge in an oil production plant.

[0039] Specifically, the Ralstonia Q6 strain achieves a 100% removal rate of pyridine at concentrations ≤400 mg / L (preferably 200–400 mg / L) within a 72-hour treatment cycle, and removal rates of ≥98.3%, ≥86.2%, and ≥73.5% for pyridine concentrations of 600 mg / L, 800 mg / L, and 1000 mg / L, respectively.

[0040] Preferably, the removal rates of Rolstonia Q6 for pyridine at concentrations of 200 mg / L, 400 mg / L, 600 mg / L, 800 mg / L, and 1000 mg / L, respectively, are 100%, 100%, 98.3%, 86.2%, and 73.5% within a 72-hour treatment cycle.

[0041] More preferably, at a pyridine concentration of 600 mg / L, the strain entered the logarithmic growth phase from 24 h and reached its maximum biomass at 48 h, confirming that the bacteria maintained an active proliferation state during the efficient degradation process.

[0042] The Roldstonia Q6 strain of the present invention is suitable for the bioremediation of industrial wastewater, groundwater and contaminated soil containing pyridine. It exhibits significant tolerance and decomposition ability to high concentrations of pyridine pollutants and can effectively decompose pyridine pollutants in actual wastewater. It has good engineering application value in the field of treating high concentration pyridine wastewater.

[0043] Another specific embodiment of the present invention discloses a pyridine-degrading formulation, wherein the formulation comprises the aforementioned Rolstonia Q6.

[0044] Another specific embodiment of the present invention discloses a method for degrading pyridine using the above-mentioned Roldstone bacteria Q6, comprising the following steps:

[0045] (1) Expand the culture of Rolstonia Q6, centrifuge to obtain a bacterial suspension;

[0046] (2) Dissolve pyridine in an inorganic salt culture medium to obtain a culture medium containing pyridine;

[0047] (3) The bacterial suspension is added to the culture medium containing pyridine and cultured to degrade the pyridine.

[0048] Specifically, in step (1), the expanded culture includes: inoculating the Ralstonia Q6 into LB medium and culturing it at 35–37°C (e.g., 35°C, 35.5°C, 36°C, 36.5°C, 37°C) and 120–150 r / min (e.g., 120 r / min, 125 r / min, 130 r / min, 135 r / min, 140 r / min, 145 r / min, 150 r / min) until the logarithmic growth phase. The bacterial cells are then collected by centrifugation at 5000–8000 r / min (5000 r / min, 5500 r / min, 6000 r / min, 6500 r / min, 7000 r / min, 7500 r / min, 8000 r / min) for 5–10 min (e.g., 5 min, 6 min, 7 min, 8 min, 9 min, 10 min). The bacterial cells are washed with sterile physiological saline and resuspended to an appropriate concentration to obtain the bacterial suspension.

[0049] Preferably, the pH value of the LB medium is 7.0.

[0050] Specifically, in step (2), the concentration of pyridine in the pyridine-containing culture medium is ≥200 mg / L, preferably 200 to 1000 mg / L, for example, 200 mg / L, 300 mg / L, 400 mg / L, 500 mg / L, 600 mg / L, 700 mg / L, 800 mg / L, 900 mg / L, 1000 mg / L.

[0051] Preferably, the pH of the pyridine-containing culture medium is 7.0.

[0052] It should be noted that the LB medium in this invention comprises: 10 g / L tryptone, 5 g / L yeast extract, and 10 g / L NaCl. The inorganic salt medium comprises: 4.26 g / L Na₂HPO₄, 2.65 g / L KH₂PO₄, 0.20 g / L MgSO₄·7H₂O, 0.006 g / L CaCl₂, and 1 mL / L trace element solution. The trace element solution comprises: 2.10 g / L FeSO₄·7H₂O, 0.02 g / L NiCl₂·6H₂O, 0.01 g / L MnCl₂·4H₂O, and 0.01 g / L ZnSO₄·7H₂O.

[0053] Specifically, in step (3), the volume of the bacterial suspension is 5% of the culture medium volume.

[0054] Specifically, in step (3), the cultivation is carried out under the conditions of 35-37℃ (e.g., 35℃, 35.5℃, 36℃, 36.5℃, 37℃) and 120-180r / min (120r / min, 125r / min, 130r / min, 135r / min, 140r / min, 145r / min, 150r / min, 155r / min, 160r / min, 165r / min, 170r / min, 175r / min, 180r / min) with light protection and shaking, and the cultivation time is 72-80h (e.g., 72h, 73h, 74h, 75h, 76h, 77h, 78h, 79h, 80h).

[0055] Another specific embodiment of the present invention discloses the application of the above-mentioned Rolstonia Q6 in the degradation of pyridine-containing wastewater.

[0056] The technical solution of the present invention will be further explained below with reference to specific embodiments.

[0057] The oilfield sludge samples in this invention were collected from an oilfield sludge sample from an oilfield production plant in Shenyang City, Liaoning Province.

[0058] Example 1

[0059] 1. Enrichment and domestication of microbial communities

[0060] (1) Collect oilfield sludge samples from oil production plants as the initial bacterial source. Take 10 g of sludge sample and inoculate it into 100 mL of inorganic salt culture medium containing 50 mg / L pyridine. Place it in a constant temperature shaking incubator and carry out light-proof shaking culture at 150 r / min under constant temperature conditions of 35℃. The inorganic salt culture medium formula includes: disodium hydrogen phosphate (Na₂HPO₄) 4.26 g / L, potassium dihydrogen phosphate (KH₂PO₄) 2.65 g / L, magnesium sulfate heptahydrate (MgSO₄·7H₂O) 0.20 g / L, and calcium chloride (CaCl₂) 0.006 g / L. The trace element solution composition includes: ferrous sulfate heptahydrate (FeSO₄·7H₂O) 2.10 g / L, nickel chloride hexahydrate (NiCl₂·6H₂O) 0.02 g / L, manganese chloride tetrahydrate (MnCl₂·4H₂O) 0.01 g / L, and zinc sulfate heptahydrate (ZnSO₄·7H₂O) 0.01 g / L. The trace element addition amount is 1 mL / L. The initial pH of the culture medium is adjusted to 7.0 with 0.1 mol / L sodium hydroxide solution.

[0061] (2) Every 14 days, 10 mL of culture medium was transferred to 90 mL of fresh culture medium, and the pyridine concentration gradient was increased in stages: the first stage was 50 mg / L for 14 days, the second stage was 200 mg / L for 14 days, the third stage was 400 mg / L for 14 days, the fourth stage was 600 mg / L for 14 days, and the fifth stage was 800 mg / L for 28 days. The entire acclimatization period lasted 84 days. At the end of the period, the removal rate of 800 mg / L pyridine by the acclimatized bacteria was detected by high performance liquid chromatography, which showed that the removal rate was 91.3%, and a stable pyridine-degrading bacterial community was formed.

[0062] 2. Screening of strains

[0063] (1) Take 1 mL of the above-mentioned domesticated and mature bacterial culture medium, add 9 mL of sterile physiological saline and perform a tenfold serial dilution, and prepare 1000-fold serial dilutions. -4 10 -5 10 -6 10 -7Four concentration gradients of bacterial suspension were prepared. 100 μL of each dilution was evenly spread onto the surface of an inorganic salt solid medium containing 300 mg / L pyridine (the medium was prepared by adding 15 g / L agar to the inorganic salt medium). The plates were sterilized by flame burning over an alcohol lamp using a sterile spreader. After cooling, the bacterial suspension was evenly spread at a 45° angle. Three replicates were prepared for each dilution. The plates were incubated upside down in a 35°C incubator and observed for 7 days.

[0064] (2) In 10 -5 On the dilution plates, round, milky-white colonies with a diameter of 2-3 mm were observed, with smooth, moist surfaces and regular edges. Thirty morphologically distinct colonies were picked using a sterile inoculation needle and purified separately on fresh inorganic salt solid medium plates containing 300 mg / L pyridine using the four-zone streak method. Each plate was incubated upside down at 35°C for 48 hours after streaking, and the purification process was repeated three times.

[0065] (3) The purified strain was inoculated into a liquid medium containing 300 mg / L pyridine and cultured with shaking at 35°C and 150 r / min for 7 days. The residual concentration of pyridine was detected by high performance liquid chromatography to obtain a strain that had completely degraded pyridine, namely Rolstonia Q6 (hereinafter referred to as strain Q6).

[0066] 3. Identification of strains

[0067] (1) The Q6 strain obtained above was inoculated into a fresh inorganic salt solid culture medium containing 300 mg / L pyridine. After culturing for 3 days, once the strain showed vigorous and uniform growth in the solid culture medium, the morphological characteristics of its colonies were observed, such as... Figure 1 As shown. The colony characteristics of strain Q6 are round, small in inorganic salt medium, with regular, slightly raised edges, and a smooth, moist surface that is easy to pick up; the colonies are translucent and white in color, with little difference in color between the front and back sides; the colonies are odorless, highly viscous, and very soft in texture.

[0068] (2) Gram staining experiment was performed on strain Q6. First, an appropriate amount of bacterial cells was picked up with an inoculation loop and spread evenly on a glass slide. The spread should be fixed. Then, the bacteria were stained with ammonium oxalate crystal violet and iodine-potassium iodide solution for 1 min, followed by washing with water. The bacteria were then destained with a neutral decolorizing agent such as ethanol, and then stained with safranin solution for 1 min. The bacteria turned red, indicating that they were Gram-negative bacteria. The staining image is shown in the figure. Figure 2 .

[0069] (3) Strain Q6 was subjected to scanning electron microscopy. At 20Kx magnification, strain Q6 appeared as a long rod shape, with an average cell size of approximately 0.2 × 1.6 μm. See the electron micrograph below. Figure 3 .

[0070] (4) Identification of 16S rDNA of strain Q6. The purified DNA of strain Q6 was extracted using a bacterial DNA extraction kit (Zymo Research, USA). The 16S rDNA of strain Q6 was obtained using universal bacterial 16S primers (upstream primer: 16S(27F)5'-AGAGTTTGATCCTGGCTCAG-3', downstream primer: 16S(1492R)5'-GGTTACCTTGTTACGACTT-3'). The 16S rDNA of strain Q6 is shown in SED ID No. 1.

[0071] See phylogenetic tree Figure 4 The results showed that the 16S rDNA sequence of strain Q6 was 1430 bp in length. Strain Q6 was most closely related to the genus Ralstonia, with a highest homology similarity of 99.79%. Therefore, strain Q6 was identified as belonging to the genus Ralstonia, specifically the phylum Proteobacteria, class Betaproteobacteria, order Burkholderiales, family Burkholderiaceae, and genus Ralstonia.

[0072] Application Example 1

[0073] Experimental verification of the pyridine degradation performance of strain Q6

[0074] (1) The Q6 strain was inoculated into LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L NaCl, pH 7.0), and cultured at 37℃ and 140 r / min for 24 h with shaking. The cells were collected by centrifugation at 5000 r / min for 5 minutes, washed with sterile physiological saline and resuspended to a concentration of OD600=1.5±0.1 to obtain the Q6 bacterial suspension.

[0075] Inorganic salt culture media (composition as in Example 1) with different pyridine concentrations (200 mg / L, 400 mg / L, 600 mg / L, 800 mg / L, and 1000 mg / L) were prepared. Q6 bacterial suspension was inoculated into each concentration system at a volume ratio of 5%. The cultures were incubated at 35°C and 150 rpm in the dark with shaking. Every 12 hours, 5 mL samples were taken, filtered through a 0.22 μm organic filter membrane, and immediately analyzed by high-performance liquid chromatography (HPLC) to determine the pyridine concentration.

[0076] Experimental data on pyridine removal rate are as follows Figure 5As shown, the results indicate that strain Q6 can completely degrade pyridine at concentrations of 200 mg / L and 400 mg / L within 72 hours. The removal rates of pyridine at concentrations of 600 mg / L, 800 mg / L, and 1000 mg / L within 72 hours are 98.3%, 86.2%, and 73.5%, respectively.

[0077] (2) Further, the degradation curve of strain Q6 with 600 mg / L pyridine as the sole carbon source is as follows: Figure 6 As shown.

[0078] according to Figure 6 It can be seen that the degradation rate increases from the 30th hour, and the removal rate of pyridine reaches 98.3% at the end of 72 hours.

[0079] For the degradation experiment of pyridine with an initial concentration of 600 mg / L, 4 ml samples were taken every 12 hours to measure the absorbance at a wavelength of 600 nm (see...). Figure 7 The results showed that the changes in bacterial biomass were synchronized with the degradation process. The bacteria entered the logarithmic growth phase at 24 hours, reached a peak growth of 0.51 at 48 hours, and maintained a growth of 0.46 at 72 hours, confirming that the bacteria maintained an active proliferation state during the efficient degradation process.

[0080] Strain Q6 entered its logarithmic growth phase at 24 hours and reached its maximum biomass at 48 hours, confirming that the bacteria maintained an active, proliferative state during the efficient degradation process. This strain exhibited significant tolerance and decomposition ability to high concentrations of pyridine pollutants, effectively decomposing pyridine pollutants in actual wastewater, and thus has good engineering application value in the treatment of high-concentration pyridine wastewater.

[0081] Application Example 2

[0082] Degradation experiments of pyridine and naphthalene by strain Q6 at different NaCl concentrations

[0083] (1) Prepare inorganic salt culture media with different salinities (0.0%, 1.0%, 2.0%, 3.0%, 4.0%, and 5.0% NaCl, respectively) (composition as in Example 1). Inoculate Q6 bacterial suspension into different salinity systems containing 600 mg / L pyridine at a volume ratio of 5%. Incubate at 35℃ and 150 r / min in the dark with shaking. After 72 h, take 5 mL of sample, filter through a 0.22 μm organic filter membrane, and immediately analyze the pyridine concentration by high performance liquid chromatography.

[0084] Pyridine degradation effect such as Figure 8 As shown, the strain maintained a 97.3% removal rate of pyridine at a salinity of 2.0%, but the removal rate decreased to 78.1% at a salinity of 5.0%.

[0085] (2) Further, the Q6 bacterial suspension was inoculated into systems containing 50 mg / L naphthalene at different salinities at a volume ratio of 5%. The cultures were incubated at 35°C and 150 r / min in the dark with shaking. After 168 h, the naphthalene concentration was determined by gas chromatography.

[0086] Naphthalene degradation effect such as Figure 9 As shown, at a salinity of 0.0%, strain Q6 achieved a degradation rate of only 50.4% for 50 mg / L naphthalene. However, as the salinity gradually increased to 5.0%, the degradation rate dropped to 8.6%, indicating almost no degradation effect.

[0087] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any changes or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention.

Claims

1. A Ralstonia sp. Q6 with accession number CGMCC No.33402.

2. The Ralstonia Q6 according to claim 1, characterized in that, The Rolstonia Q6 strain contains the 16S rDNA shown in SEQ ID No.

1.

3. The Roldstone bacteria Q6 according to claim 1, characterized in that, The aforementioned Rolstonia Q6 was isolated from oilfield sludge in an oil production plant.

4. The Ralstonia Q6 according to any one of claims 1-3, characterized in that, The Ralstonia Q6 strain achieved a 100% removal rate of pyridine at concentrations ≤400 mg / L within a 72-hour treatment cycle, and removal rates of ≥98.3%, ≥86.2%, and ≥73.5% for pyridine at concentrations of 600 mg / L, 800 mg / L, and 1000 mg / L, respectively.

5. A formulation for degrading pyridine, characterized in that, Includes Rolstonia Q6 as described in any one of claims 1-4.

6. A method for degrading pyridine using Rolstonia Q6 according to any one of claims 1-4, characterized in that, Includes the following steps: (1) Expand the culture of Rolstonia Q6, centrifuge to obtain a bacterial suspension; (2) Dissolve pyridine in an inorganic salt culture medium to obtain a culture medium containing pyridine; (3) The bacterial suspension is added to the culture medium containing pyridine and cultured to degrade the pyridine.

7. The method according to claim 6, characterized in that, In step (1), the expansion culture specifically includes: inoculating the Ralstonia Q6 into LB medium, culturing it at 35-37°C and 120-150 r / min to the logarithmic growth phase, then collecting the bacterial cells by centrifugation at 5000-8000 r / min for 5-10 min, washing the bacterial cells with sterile physiological saline and resuspending them to an appropriate concentration to obtain the bacterial suspension.

8. The method according to claim 6, characterized in that, In step (2), the concentration of pyridine in the pyridine-containing culture medium is ≥200 mg / L.

9. The method according to claim 6, characterized in that, In step (3), the culture is carried out under the conditions of 35-37℃ and 120-180r / min in the dark with shaking, and the culture time is 72-80h.

10. The use of Rolstonia Q6 according to any one of claims 1-4 in the degradation of pyridine-containing wastewater.

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