5-fluorouracil degrading bacteria yj07 and application thereof

Abstract

This invention relates to the field of microbial technology, and proposes a 5-fluorouracil-degrading bacterium YJ07 and its applications. The 5-fluorouracil-degrading bacterium YJ07 is *Agromycetes mirifica* (…). Agromyces mediolanus YJ07 was deposited on December 18, 2025, 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. 1.65195. The above technical solution addresses the problems of poor tolerance and low degradation efficiency of strains to 5-fluorouracil in related technologies.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, specifically to a 5-fluorouracil-degrading bacterium YJ07 and its applications. Background Technology

[0002] 5-Fluorouracil (5-FU) is a first-line chemotherapy drug widely used in the treatment of various malignant tumors, including colorectal cancer and breast cancer. It is a fluorine-containing organic compound with genotoxic and reproductive toxic effects. Furthermore, its CF bond structure is stable and difficult to degrade in the natural environment. This drug cannot be completely metabolized in the human body, and large quantities are excreted in urine and other forms, either unchanged or as metabolites. Once it enters wastewater systems, it poses a potential risk to aquatic life.

[0003] Currently, the main methods for degrading 5-fluorouracil are chemical catalytic degradation. These methods have drawbacks such as harsh reaction conditions, high costs, and potential secondary pollution. Biodegradation, on the other hand, has become a research hotspot due to its advantages of high efficiency, environmental friendliness, and mildness. However, research on specific degrading bacteria for 5-fluorouracil is still incomplete. Existing strains have insufficient degradation efficiency and specificity for 5-fluorouracil, and most strains have poor tolerance to 5-fluorouracil, making it difficult for them to survive stably and perform degradation functions in environments containing 5-fluorouracil, thus limiting their practical application. Summary of the Invention

[0004] This invention proposes a 5-fluorouracil-degrading bacterium YJ07 and its application, which solves the problems of poor tolerance to 5-fluorouracil and low degradation efficiency of strains in related technologies.

[0005] The technical solution of the present invention is as follows: This invention proposes a 5-fluorouracil-degrading bacterium YJ07, wherein the 5-fluorouracil-degrading bacterium YJ07 is Agrobacterium mirificum ( Agromyces mediolanus YJ07 was deposited on December 18, 2025, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, with accession number CGMCCNo.1.65195.

[0006] As a further technical solution, the culture temperature during the cultivation of the 5-fluorouracil-degrading bacteria is 30°C.

[0007] As a further technical solution, the pH of the culture medium is 6 during the cultivation of the 5-fluorouracil degrading bacteria.

[0008] As a further technical solution, the 5-fluorouracil degrading bacterium YJ07 has a tolerance concentration of 5-fluorouracil of 50 μg / mL. In a 5-fluorouracil system with this concentration, the growth of the 5-fluorouracil degrading bacterium YJ07 is not affected.

[0009] This invention also proposes the application of a 5-fluorouracil-degrading bacterium, YJ07, in the degradation of 5-fluorouracil.

[0010] As a further technical solution, the application of the 5-fluorouracil degrading bacterium YJ07 in the preparation of 5-fluorouracil degrading products.

[0011] As a further technical solution, the application of the 5-fluorouracil degrading bacterium YJ07 in the treatment of environmental 5-fluorouracil pollution.

[0012] As a further technical solution, the 5-fluorouracil degrading bacterium YJ07 is used in the preparation of products for treating environmental 5-fluorouracil pollution.

[0013] The present invention also proposes a microbial agent containing the 5-fluorouracil degrading bacterium YJ07 and / or metabolites of the 5-fluorouracil degrading bacterium YJ07.

[0014] As a further technical solution, the bacterial agent has the property of degrading 5-fluorouracil.

[0015] The working principle and beneficial effects of this invention are as follows: 1. The 5-fluorouracil degrading bacterium YJ07 in this invention has high tolerance to 5-fluorouracil. In a system with a 5-fluorouracil concentration of 50 μg / mL, the growth of the 5-fluorouracil degrading bacterium YJ07 is not affected.

[0016] 2. The 5-fluorouracil degrading bacterium YJ07 in this invention has good degradability of 5-fluorouracil. When a 250mL Erlenmeyer flask is used as the culture container, and 50mL of degradation medium with pH 6 is added to the Erlenmeyer flask, under the conditions of culture temperature of 30℃ and inoculum amount of 5-fluorouracil degrading bacterium YJ07 of 1%, when the initial concentration of 5-fluorouracil is 0.5~1μg / mL, the 5-fluorouracil degrading bacterium YJ07 can almost completely degrade 5-fluorouracil within 48h. Attached Figure Description

[0017] The present invention will now be described in further detail with reference to the accompanying drawings and specific embodiments.

[0018] Figure 1 The results of the screening test for 5-fluorouracil degrading strains are shown in the figure. A is the blank control group, B is the experimental group with only YJ07 added, C is the experimental group with only YJ08 added, and D is the experimental group with a mixture of YJ07 and YJ08. Figure 2 The degradation of 5-fluorouracil by the 5-fluorouracil-degrading bacterium YJ07 over 6 days was shown. Figure 3 The results of the catalase test for 5-fluorouracil degrading bacteria YJ07 are shown in the figure. The left figure is YJ07 without hydrogen peroxide solution treatment, and the right figure is the blank group with only hydrogen peroxide solution added and YJ07 after hydrogen peroxide solution treatment. Figure 4 The results of the oxidase test of 5-fluorouracil degrading bacteria YJ07 are shown in the figure. The left figure is YJ07 before treatment with 1% dimethyl p-phenylenediamine hydrochloride solution, and the right figure is YJ07 after treatment with 1% dimethyl p-phenylenediamine hydrochloride solution. Figure 5 The results of the methyl red test for 5-fluorouracil degrading bacteria YJ07 are shown in the figure. The left figure shows the blank group, YJ07, Escherichia coli, and Bacillus subtilis before the addition of methyl red reagent, and the right figure shows the blank group, YJ07, Escherichia coli, and Bacillus subtilis after the addition of methyl red reagent. Figure 6 The results of the Voges-Pyr test for 5-fluorouracil-degrading bacteria YJ07 are shown in the figure. The left figure shows the blank group, YJ07, Escherichia coli, and Bacillus subtilis before treatment with α-naphthol alcohol solution and potassium hydroxide solution. The right figure shows the blank group, YJ07, Escherichia coli, and Bacillus subtilis after treatment with α-naphthol alcohol solution and potassium hydroxide solution. Figure 7 The results of PCR amplification of 16S rDNA from the 5-fluorouracil-degrading bacterium YJ07 are shown. Figure 8 The effect of different liquid volumes on the degradation of 5-fluorouracil by the 5-fluorouracil-degrading bacterium YJ07; Figure 9 The effect of different inoculum amounts on the degradation of 5-fluorouracil by the 5-fluorouracil-degrading bacterium YJ07; Figure 10 The effect of different pH values ​​on the degradation of 5-fluorouracil by the 5-fluorouracil-degrading bacterium YJ07; Figure 11 The effect of different temperatures on the degradation of 5-fluorouracil by the 5-fluorouracil-degrading bacterium YJ07.

[0019] Instructions for the Preservation of Biological Materials The 5-fluorouracil-degrading bacterium YJ07 provided by this invention is Agrobacterium mirifica ( Agromyces mediolanus YJ07 was deposited on December 18, 2025, 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. 1.65195. Detailed Implementation

[0020] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0021] In the following embodiments, LB liquid medium consists of the following components: 10g tryptone, 5g yeast extract, 10g NaCl, water added to 1L, pH=7.2; LB solid medium consists of the following components: 10g tryptone, 5g yeast extract, 10g NaCl, 20g agar, and water added to 1L, pH=7.2; The bacterial screening liquid culture medium consists of the following components: 10.0 g tryptone, 1.5 g Na2HPO4, 0.5 g KH2PO4, 1.0 g NaCl, 0.2 g MgSO4·7H2O, 10 mL trace element solution, and water added to 1 L, pH=7.2; The solid culture medium for bacterial screening consists of the following components: 10.0 g tryptone, 1.5 g Na2HPO4, 0.5 g KH2PO4, 1.0 g NaCl, 0.2 g MgSO4·7H2O, 20 g agar, 10 mL trace element solution, and water to a final volume of 1 L, pH=7.2. The trace element solution comprises the following components: MnCl•4H2O 0.223g; ZnSO4•7H2O 0.086g; H3BO3 0.062g; KI 0.0083g; Na2MoO4•2H2O 0.0025g; CuSO4•5H2O 0.0003g; CoCl2•6H2O 0.0003g, dissolved in water and brought to a final volume of 100mL.

[0022] Example 1 1.1 Enrichment and domestication of degrading strains Soil from the grassland of Hebei University of Science and Technology (address: No. 26 Yuxiang Street, Yuhua District, Shijiazhuang City, Hebei Province) was selected, placed in a container, and irrigated with 200 mL of 5-fluorouracil aqueous solution containing 400 mg / L every week for 2 months to obtain 5-fluorouracil contaminated soil. Weigh 3g of 5-fluorouracil-contaminated soil and place it in a 250mL Erlenmeyer flask containing 100mL of sterile water. Shake for 30min at 30℃ and 160rpm. Take the supernatant and inoculate it into LB liquid medium at a 5% inoculum. Incubate at 30℃ and 160rpm for 24h with shaking. After standing, take the supernatant. The supernatant was inoculated at 5% into 50 mL of LB liquid medium with a 5-fluorouracil concentration of 10 mg / L. After culturing in the dark for 5 days at 30°C and 160 rpm in a shaker, the supernatant was then transferred to LB liquid medium with 5-fluorouracil concentrations of 20 mg / L, 30 mg / L, 40 mg / L, and 50 mg / L for gradient acclimatization culture, following the same culturing method. Take 1 mL of the culture medium obtained from the last domestication and prepare it into a 1000 dilution. -1 10 -2 and 10 -3 100 μL of the bacterial suspension was spread onto a bacterial screening solid medium plate with a 50 mg / L concentration of 5-fluorouracil and incubated at 30°C and 60% RH for 5 days. Colonies of different shapes, sizes and colors were repeatedly streaked and purified. Finally, two strains were isolated and named YJ07 and YJ08, respectively.

[0023] 1.2 Screening of degrading strains The pH of the bacterial screening liquid culture medium was adjusted to 7 with 1 mol / L NaOH. Then, 100 mL of the bacterial screening liquid culture medium was placed into 250 mL Erlenmeyer flasks and sterilized at 121 °C for 20 min. 5 mg / mL of 5-fluorouracil solution was added to each Erlenmeyer flask to make the initial concentration of 5-fluorouracil in each flask 1.0 ± 0.2 μg / mL. The experiment was set up with 4 groups (including 1 blank control group and 3 experimental groups), with 3 parallel samples in each group: the blank control group was not inoculated with YJ07 and YJ08; the 3 experimental groups were YJ07 group with 1% inoculation, YJ08 group with 1% inoculation, and YJ07 and YJ08 mixed inoculation group (YJ07 and YJ08 were mixed in equal proportions, with an inoculation amount of 1%). They were then placed in a shaker at 30℃ and 160rpm for 6 days to degrade. Samples were taken on day 0 and day 6, and centrifuged at 4℃ and 12000rpm for 10min. The supernatant was collected and repeated twice. The samples were stored at -20℃ for later testing. Take 20 μL of a 5 mg / mL 5-fluorouracil solution and add it to 9980 μL of sterile water to prepare a 10 μg / mL 5-fluorouracil solution. Then, using the 10 μg / mL 5-fluorouracil solution as a stock solution, dilute it with bacterial screening liquid medium to prepare standard samples with concentrations of 0.005, 0.01, 0.02, 0.05, 0.1, 0.2, 0.5, 1, 1.2, and 1.5 μg / mL. The concentration of 5-fluorouracil in the sample was determined using liquid chromatography-mass spectrometry (LC-MS). The chromatographic column was a C18 reversed-phase column (2.1 × 150 mm, 1.8 μm, Agilent Technologies). The mobile phase consisted of 0.1% formic acid and methanol. The injection volume was 0.5 μL, the flow rate was 0.2 mL / min, and the analysis time was 10 min. For the first 3 min, isocratic elution was performed using a 5% methanol to 95% formic acid ratio. For the subsequent 7 min, a gradient elution was used, with the methanol content in the mobile phase linearly increasing from 5% to 95%. Mass spectrometry was performed using an electrospray ionization (ESI) source in negative ion mode. The spray voltage was 1.5 kV, and the capillary temperature was 300 °C. Data acquisition was performed in MRM mode. The precursor ion was 129.1 m / z, and the daughter ions were 59.1 and 42.0 m / z, with fragmentation voltages of 80 V and collision voltages of 20 V and 35 V, respectively. Test results are as follows Figure 1 As shown, the results indicate that 5-fluorouracil itself did not undergo hydrolysis. On day 6, YJ07 completely degraded 5-fluorouracil, but YJ08 and the mixture of YJ07 and YJ08 did not completely degrade 5-FU.

[0024] Furthermore, without changing the degradation test method, only YJ07 was used to degrade 5-fluorouracil for 6 days. Samples were taken every 24 hours during the experiment, centrifuged at 4℃ and 12000 rpm for 10 min, and the supernatant was collected. This process was repeated twice and stored at -20℃ for analysis. Finally, quantitative analysis of the samples was performed using liquid chromatography-mass spectrometry (LC-MS), and the results are as follows: Figure 2 As shown, YJ07 completely degraded 5-fluorouracil on the second day.

[0025] 1.3 Physiological and biochemical identification of degrading strains 1.3.1 Catalase test YJ07 was incubated at 30℃ and 60% RH for 48 hours. Then, a single colony was picked up using a sterile disposable plastic inoculation loop and smeared onto a glass slide. A 3% hydrogen peroxide solution was then added to completely cover the bacterial cells. The presence of persistent bubbles within 30 seconds indicated a positive result, while the absence of bubbles indicated a negative result. The test results are as follows: Figure 3 As shown, YJ07 produces bubbles, indicating that YJ07 produces catalase during its growth process.

[0026] 1.3.2 Oxidase test A single colony of YJ07 cultured for 48 hours was picked using a sterile disposable plastic inoculation loop and spread onto a filter paper strip. Then, a 1% dimethyl-p-phenylenediamine hydrochloride solution was added to completely cover the colony. A positive result was indicated by the colony turning purple or blue within 10 seconds, and a negative result was indicated by no color change within 2 minutes. The test results are as follows: Figure 4As shown, YJ07 showed no color change, indicating that YJ07 did not produce cytochrome C oxidase.

[0027] 1.3.3 Methyl Red (MR) Test YJ07, positive control *Escherichia coli*, and negative control *Bacillus subtilis* were inoculated into test tubes containing 6 mL of glucose peptone liquid medium, respectively. The cultures were incubated statically at 30°C and 60% RH for 48 h. The bacterial cultures were then mixed thoroughly, and 3 mL was added to new test tubes. Three drops of methyl red reagent were added to each tube, and the color change was immediately observed. The experimental results are as follows: Figure 5 As shown, YJ07 is orange-yellow in color, and the result is negative.

[0028] 1.3.4 Volp II (VP) Test YJ07 bacteria cultured for 48 hours in test tubes containing 6 mL of glucose peptone broth, *E. coli* (negative control), and *Bacillus subtilis* (positive control) were mixed thoroughly. 2.5 mL of each mixture was added to a new test tube. Then, 0.6 mL of 6% α-naphthol ethanol solution was added, followed by 0.2 mL of 40% potassium hydroxide solution. The mixture was shaken for 5 minutes, and the color change was observed. The experimental results are as follows: Figure 6 As shown, YJ07 is yellow, indicating a negative result.

[0029] 1.4 Gene identification of degrading strains 1.4.1 16S rDNA assay Total DNA was extracted from YJ07 according to the Ezup column-based genomic DNA extraction kit. Using the extracted total DNA as a template, and 27F and 1492R as amplification primers, the 16S rDNA of YJ07 was amplified. The obtained PCR products were detected by electrophoresis on a 1.5% agarose gel. The results are shown below. Figure 7 As shown, after the amplification results were confirmed, the sample was sent to a testing institution for sequencing, and a gene sequence of 1458 bp was obtained after sequencing.

[0030] 1.4.2 Phylogenetic Analysis The obtained 16S rDNA sequences were submitted to the National Center for Biotechnology Information (NCBI) for alignment analysis using BLAST to obtain highly homologous gene sequences. A phylogenetic tree was then constructed using MEGA 12 software with a neighbor-joining method. The results showed that YJ07 and... Agromyces mediolanus The samples showed a homology of over 99%. Based on morphological characteristics, physiological and biochemical characteristics, and 16S rDNA genetic development analysis, YJ07 was identified as Agrobacterium mirificum.

[0031] Example 2 1. Scale-up culture and washing of degrading bacteria YJ07 YJ07 cells grown on LB solid medium were picked and inoculated into 50 mL of LB liquid medium and cultured in a shaker at 160 rpm and 30 °C for 24 h to obtain the expanded culture solution. Transfer 20 mL of the expanded bacterial culture to a 50 mL centrifuge tube and centrifuge at 6000 rpm for 10 min at room temperature, discarding the supernatant. Then add 20 mL of PBS buffer, gently disperse the bacteria in the buffer, centrifuge at 6000 rpm for 10 min at room temperature, discard the supernatant, repeat the washing process three times, and then dissolve the bacteria in degradation medium, adjusting the OD... 600 It is 1.0.

[0032] 2. Degradation characteristics of degrading bacteria YJ07 2.1 Determination of Initial Liquid Volume The pH of the bacterial screening liquid culture medium was adjusted to 7 with 1 mol / L NaOH. Then, 25 mL, 50 mL, 75 mL, 100 mL, and 125 mL of the bacterial screening liquid culture medium were respectively placed into 250 mL Erlenmeyer flasks and sterilized at 121 °C for 20 min. 1 mg / mL of 5-fluorouracil solution was added to each Erlenmeyer flask to make the initial concentration of 5-fluorouracil in each flask 1.0 ± 0.2 μg / mL. The experiment was set up with 10 groups (including 5 blank control groups and 5 experimental groups), with 3 parallel samples in each group: the blank control group was not inoculated with YJ07; the inoculation amount of YJ07 in the 5 experimental groups was 1%, corresponding to liquid volumes of 25 mL, 50 mL, 75 mL, 100 mL and 125 mL respectively. They were then placed in a shaker at 30℃ and 160 rpm for 48 h of degradation. Samples were taken at 0 h and 48 h and centrifuged at 4℃ and 12000 rpm for 10 min. The supernatant was collected and repeated twice. The samples were stored at -20℃ for analysis. Experimental results are as follows Figure 8 As shown, the optimal liquid volume is 50 mL. Under these conditions, there is sufficient oxygen, which can promote better growth and degradation of the strain, with a degradation amount of 1.10 μg / mL.

[0033] 2.2 Determination of initial inoculation dose Adjust the pH of the bacterial screening liquid culture medium to 7 with 1 mol / L NaOH, then take 50 mL of the bacterial screening liquid culture medium and put it into 250 mL Erlenmeyer flasks. Sterilize at 121℃ for 20 min. Add 1 mg / mL of 5-fluorouracil solution to each Erlenmeyer flask to make the initial concentration of 5-fluorouracil in each flask 1 μg / mL. The experiment was set up with 10 groups (including 5 blank control groups and 5 experimental groups), with 3 parallel samples in each group: the blank control group was not inoculated with YJ07; the inoculation amount of YJ07 in the 5 experimental groups was 0.5%, 1%, 3%, 5%, and 7%, respectively. They were then placed in a shaker at 30℃ and 160 rpm for 48 h to degrade. Samples were taken at 0 h and 48 h, and centrifuged at 4℃ and 12000 rpm for 10 min. The supernatant was collected and repeated twice. The samples were stored at -20℃ for later analysis. Experimental results are as follows Figure 9 As shown, when the inoculum amount of YJ07 is 1%~7%, the degradation effect on 5-fluorouracil is stable, and the degradation amount is basically maintained at 1.10-1.11 μg / mL, which is almost complete degradation.

[0034] 2.3 Determination of initial pH The bacterial screening liquid culture medium was adjusted to pH 5, 6, 7, 8 and 9 respectively with 1 mol / L NaOH and 1 mol / L HCl. Then, 50 mL of the bacterial screening liquid culture medium was placed into 250 mL Erlenmeyer flasks and sterilized at 121 °C for 20 min. 1 mg / mL of 5-fluorouracil solution was added to each Erlenmeyer flask to make the initial concentration of 5-fluorouracil in each flask 1.0 ± 0.2 μg / mL. The experiment was set up with 10 groups (including 5 blank control groups and 5 experimental groups), with 3 replicates in each group: the blank control group was not inoculated with YJ07; the inoculation amount of YJ07 in the 5 experimental groups was 1%, corresponding to pH 5, 6, 7, 8 and 9 respectively. They were then placed in a shaker at 30℃ and 160rpm for 48h of degradation. Samples were taken every 8h and centrifuged at 4℃ and 12000rpm for 10min. The supernatant was collected and repeated twice. The samples were stored at -20℃ for analysis. Experimental results are as follows Figure 10 As shown, YJ07 exhibited the best degradation effect on 5-fluorouracil at pH 6, with a degradation amount reaching 1.03 μg / mL after 48 h, which was almost complete degradation; no degradation was observed at pH 5; and although degradation occurred at pH 7-9, both the rate and amount of degradation were lower than at pH 6.

[0035] 2.4 Determination of Initial Temperature The bacterial screening liquid culture medium was adjusted to pH 6 with 1 mol / L HCl. Then, 50 mL of the bacterial screening liquid culture medium was placed into 250 mL Erlenmeyer flasks and sterilized at 121 °C for 20 min. 1 mg / mL of 5-fluorouracil solution was added to each Erlenmeyer flask to make the initial concentration of 5-fluorouracil in each flask 1.0 ± 0.2 μg / mL. The experiment was set up with 10 groups (including 5 blank control groups and 5 experimental groups), with 3 parallel samples in each group: the blank control group was not inoculated with YJ07; the inoculation amount of YJ07 in the 5 experimental groups was 1%, corresponding to the culture temperature of 20℃, 25℃, 30℃, 35℃ and 40℃ respectively. The samples were cultured in a shaker at 160 rpm for 48 h, and samples were taken every 8 h. The samples were centrifuged at 12000 rpm at 4℃ for 10 min, and the supernatant was collected. The process was repeated twice and stored at -20℃ for analysis. Experimental results are as follows Figure 11 As shown, YJ07 exhibited the best degradation effect on 5-fluorouracil at 30℃, with a degradation amount of 1.12 μg / mL after 48 hours, indicating near-complete degradation. At 20℃ and 25℃, the strain had a longer lag phase, with a slower degradation rate in the early stage, but the degradation accelerated with increasing bacterial count in the later stage, also reaching near-complete degradation after 48 hours, with degradation amounts of 1.12 μg / mL and 1.02 μg / mL, respectively. The degradation amount decreased significantly at 35℃, and there was no degradation effect at 40℃, but the strain could still grow normally. It is speculated that high temperature leads to the inactivation of degradation-related enzymes or bacterial cell damage, thus affecting the degradation effect.

[0036] 2.5 Determination of Initial Drug Concentration The bacterial screening liquid culture medium was adjusted to pH 6 with 1 mol / L HCl. Then, 50 mL of the bacterial screening liquid culture medium was placed into 250 mL Erlenmeyer flasks and sterilized at 121 °C for 20 min. 1 mg / mL of 5-fluorouracil solution was added to each Erlenmeyer flask, and the initial concentrations of 5-fluorouracil in each flask were 0.5, 1, 10, 25, and 50 μg / mL, respectively. Ten groups were set up in the experiment (including 5 blank control groups and 5 experimental groups), with 3 replicates in each group: the blank control group was not inoculated with YJ07; the inoculation amount of YJ07 in the 5 experimental groups was 1%, corresponding to initial concentrations of 5-fluorouracil of 0.5, 1, 10, 25, and 50 μg / mL, respectively. They were then incubated in a shaker at 30℃ and 160 rpm for 48 h for degradation. Samples were taken every 8 h and centrifuged at 4℃ and 12000 rpm for 10 min. The supernatant was collected, and the process was repeated twice. The samples were stored at -20℃ for analysis. The experimental results are shown in Table 1 below (rounded to three decimal places). Table 1. Effects of different 5-fluorouracil concentrations on the degradation of 5-fluorouracil by the 5-fluorouracil-degrading bacterium YJ07.

[0037] The results in Table 1 show that, under the optimal conditions optimized in the early stage, YJ07 can achieve near-complete degradation of 5-fluorouracil within 48 hours when the concentration of 5-fluorouracil is 0.5 μg / mL and 1 μg / mL. At the same time, YJ07 can tolerate a concentration of 5-fluorouracil up to 50 μg / mL, and its growth is not affected in the 5-fluorouracil system at this concentration.

[0038] The above are merely preferred 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.

Claims

1. A 5-fluorouracil-degrading bacterium, YJ07, characterized in that, The 5-fluorouracil degrading bacteria YJ07 is Lecanicillium muscarium (Gams) Agromyces mediolanus ) YJ07, which was preserved at the China General Microbiological Culture Collection Center on December 18, 2025, at 1st Yard, North Chengxilu, Chaoyang District, Beijing, with a preservation number of CGMCC No. 1.65195.

2. The 5-fluorouracil-degrading bacterium YJ07 according to claim 1, characterized in that, The culture temperature for the 5-fluorouracil-degrading bacteria is 30°C.

3. The 5-fluorouracil-degrading bacterium YJ07 according to claim 1, characterized in that, During the cultivation of the 5-fluorouracil-degrading bacteria, the pH of the culture medium was 6.

4. The 5-fluorouracil-degrading bacterium YJ07 according to claim 1, characterized in that, The 5-fluorouracil-degrading bacterium YJ07 tolerates a 50 μg / mL concentration of 5-fluorouracil, and its growth is unaffected in a 5-fluorouracil system at this concentration.

5. The application of a 5-fluorouracil-degrading bacterium, YJ07, in the degradation of 5-fluorouracil, characterized in that, The 5-fluorouracil degrading bacterium YJ07 includes the 5-fluorouracil degrading bacterium YJ07 as described in any one of claims 1 to 4.

6. The application according to claim 5, characterized in that, Application of the 5-fluorouracil degrading bacterium YJ07 in the preparation of 5-fluorouracil degrading products.

7. The application according to claim 5, characterized in that, Application of the 5-fluorouracil degrading bacterium YJ07 in the treatment of environmental 5-fluorouracil pollution.

8. The application according to claim 5, characterized in that, The application of the 5-fluorouracil degrading bacterium YJ07 in the preparation of products for treating environmental 5-fluorouracil pollution.

9. A microbial agent, characterized in that, The microbial agent contains the 5-fluorouracil degrading bacterium YJ07 as described in claims 1 to 4 and / or the metabolites of the 5-fluorouracil degrading bacterium YJ07 as described in any one of claims 1 to 4.

10. The microbial agent according to claim 9, characterized in that, The bacterial agent has the property of degrading 5-fluorouracil.

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