Aflatoxin B1 degrading bacterium and application thereof

Abstract

The invention discloses an aflatoxin B1 degrading bacterium and application thereof, and belongs to the technical field of biodegradation. The name of the degrading bacterium is bacillus paramycoides HS-1, and the preservation number of the degrading bacterium is GDMCC No: 66506. The invention further discloses a preparation method of the degrading bacterium. After the strain is cultured in an LB broth culture medium containing 2 [mu] g / mL of AFB1 for 16 h, the degradation rate of the AFB1 can reach 35% or above. The heat stability of the fermentation supernatant filtrate of the HS-1 strain for degrading the AFB1 is good, after the fermentation supernatant filtrate is treated in a boiling water bath at the temperature of 100 DEG C for 20 min, the 48-hour degradation rate of 2 microgram / mL AFB1 can be stabilized to be 70% or above, and 2 microgram / mL AFB1 can be completely degraded at the temperature of 70 DEG C, the pH value of 11 and 48 hours.

Description

Technical Field

[0001] The present invention relates to the technical field of biodegradation, and in particular to an aflatoxin B1-degrading bacterium and application thereof. Background Art

[0002] Aflatoxin B1 (AFB1) is a common mycotoxin that is widely present in grain and oil agricultural products (such as corn and peanuts) contaminated by Aspergillus flavus and Aspergillus parasiticus. Long-term exposure to AFB1 in humans can lead to various consequences, such as malignant tumors, impaired immune function, developmental delay, and complications related to malnutrition. AFB1 poisoning in animals can manifest as depression, weight loss, liver damage, and gastrointestinal bleeding, and in severe cases, death. It is classified as a Class I carcinogen by the World Health Organization's International Agency for Research on Cancer. It not only causes direct economic losses to countries around the world, but also poses serious health hazards to humans and livestock.

[0003] Due to the serious harm caused by AFB1 and its geographical distribution characteristics, international organizations and various countries and regions around the world have set strict limits on the content of AFB1 in agricultural products and foods. [13,28 The Codex Alimentarius Commission (CAC) requires a limit of 15 μg / kg for AFB1 in directly edible agricultural products, such as peanuts and almonds, and 20 μg / kg for further processed raw materials (such as corn and cottonseed). my country's latest national food safety standard, GB 2761-2017, "Limits of Mycotoxins in Food," sets a maximum limit of 20 μg / kg for AFB1 in food.

[0004] To ensure consumer safety, a variety of AFB1 detoxification methods have been developed and explored to reduce AFB1 levels in contaminated agricultural products. However, commonly used physical and chemical methods, such as high-temperature treatment, adsorption, radiation, ammonia treatment, ozone, and chlorine treatment, are often unstable and costly, and may lead to the loss of nutrients in agricultural products, the production of toxic substances, and environmental harm. In contrast, biological detoxification technologies specifically degrade AFB1 through microbial metabolism or enzymatic catalysis, offering significant advantages such as environmental friendliness, mild reaction conditions, and no degradation of the physical and chemical properties of the substrate.

[0005] Studies have shown that Bacillus, Acinetobacter and some fungi can convert AFB1 into low-toxic or non-toxic products by secreting extracellular enzymes such as laccase and peroxidase.

[71] However, existing degradation strains still have problems such as poor environmental adaptability (such as pH and temperature sensitivity) and unclear degradation pathways, which restrict their industrial application. Therefore, there is an urgent need for a strain that can efficiently degrade AFB1 at high temperatures to promote the further development and application of AFB1 biodegradation technology. Summary of the Invention

[0006] To overcome the shortcomings and deficiencies of existing microbial degradation of AFB1, the present invention provides an aflatoxin B1-degrading bacterium. This bacterium was isolated from soil contaminated with aflatoxin B1, and its fermentation supernatant filtrate exhibits significant AFB1-degrading ability. High-temperature treatment significantly enhances the AFB1-degrading ability of the fermentation supernatant filtrate.

[0007] Another object of the present invention is to provide the application of the above-mentioned aflatoxin B1-degrading bacteria.

[0008] The purpose of the present invention is achieved through the following technical solutions:

[0009] The present invention provides an aflatoxin B1-degrading bacterium named Bacillus paramycoides HS-1, which is selected from AFB1-contaminated soil in peanut-growing farmland of Guangdong Academy of Agricultural Sciences and is obtained through cultivation in an AFB1-containing enrichment medium, pressure screening, separation, and purification.

[0010] The deposit information of the Bacillus paramycoides HS-1 is as follows: depository unit: Guangdong Microbial Culture Collection Center (GDMCC), deposit date: June 13, 2025, deposit address: Institute of Microbiology, Guangdong Academy of Sciences, 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, deposit number: GDMCC No: 66506.

[0011] The 16S rDNA of the HS-1 strain has a 100% homology to that of Bacillus paramycoides. The 16S rDNA sequence of the strain is shown in SEQ ID NO: 1.

[0012] The present invention also provides an aflatoxin B1 degradation reagent, comprising the above-mentioned Bacillus mycoides HS-1, or at least one of its fermentation broth, fermentation broth supernatant, and fermentation supernatant filtrate.

[0013] Furthermore, the fermentation broth is prepared by liquid fermentation of the above-mentioned Bacillus mycoides HS-1, preferably comprising the following steps: inoculating the seed liquid of the above-mentioned Bacillus mycoides HS-1 into NB broth culture medium for fermentation, thereby obtaining the fermentation broth.

[0014] Preferably, the inoculation amount of the seed solution is 5-10% (further 5%).

[0015] Preferably, the fermentation culture conditions are 28-30° C., 180-200 r / min for 24-72 h; further 30° C., 180 rpm for 24 h.

[0016] Preferably, the seed solution is obtained by first activating Bacillus mycoides HS-1 and then picking a single colony and culturing it in NB broth medium.

[0017] The culture conditions are 28-30° C., 180-200 r / min, culturing for 10-14 h; further 30° C., 180 r / min, culturing for 12 h.

[0018] Furthermore, the fermentation broth supernatant is prepared by centrifuging the fermentation broth.

[0019] Preferably, the centrifugation conditions are 2-8° C., 8000-10000 rpm, and 8-10 min; more preferably, 4° C., 10000 rpm, and 10 min.

[0020] Furthermore, the fermentation broth supernatant filtrate is prepared by filtering the fermentation broth supernatant.

[0021] Preferably, the filtration is performed using a 0.22 μm sterile filter membrane.

[0022] The present invention also provides the use of the aflatoxin B1 degrading bacteria in degrading aflatoxin B1.

[0023] The present invention also provides the use of the aflatoxin B1 degradation reagent in degrading aflatoxin B1.

[0024] Furthermore, the use of the aflatoxin B1-degrading bacteria or degradation reagent in degrading aflatoxin B1 comprises the following steps:

[0025] (1) inoculating Bacillus paramycoides HS-1 into LB liquid medium containing AFB1 and culturing; or,

[0026] (2) inoculating the seed liquid of Bacillus paramycoides HS-1 into NB broth medium, and after fermentation, centrifuging the culture liquid, collecting the fermentation liquid supernatant, and filtering the fermentation liquid supernatant to obtain a fermentation supernatant filtrate;

[0027] (3) adding AFB1 to the fermentation supernatant filtrate of step (2) and reacting;

[0028] (4) The fermentation supernatant filtrate of step (2) was heated, cooled to room temperature, and then AFB1 was added and reacted.

[0029] Preferably, in step (1),

[0030] The final concentration of AFB1 is 0-10 μg / mL and is not 0; further, it is 2-10 μg / mL.

[0031] The culture conditions are 28-30° C., 180-200 rpm, and 16-48 h; further 30° C., 180 rpm, and 16 h;

[0032] Preferably, in step (2),

[0033] The inoculation amount of the seed liquid is 5-10% (or preferably 5%).

[0034] The fermentation culture conditions are 28-30° C., 180-200 r / min, culturing for 24-72 hours; further 30° C., 180 rpm, culturing for 24 hours.

[0035] The centrifugal conditions are 2-8° C., 8000-10000 rpm, and 8-10 min; more preferably, 4° C., 10000 rpm, and 10 min.

[0036] The filtration is performed using a 0.22 μm sterile filter membrane.

[0037] Preferably, in steps (3) and (4),

[0038] The final concentration of AFB1 is 0-10 μg / mL and is not 0; further, it is 2-10 μg / mL.

[0039] The reaction conditions are 28-30° C., 180-200 r / min, reaction for 48-72 hours; further 30° C., 180 rpm, reaction for 48 hours.

[0040] Preferably, in step (4), the heating treatment is performed at 95-100° C. for 15-20 min; further at 100° C. for 20 min.

[0041] The present invention has the following advantages and effects compared to the prior art:

[0042] (1) The present invention separates and purifies the Bacillus paramycoides HS-1 strain, which has good AFB1 degradation performance. After culturing in LB broth containing 2 μg / mL AFB1 for 16 hours, the AFB1 degradation rate can reach more than 35%.

[0043] (2) The fermentation supernatant filtrate of the Bacillus paramycoides HS-1 strain of the present invention exhibited excellent thermal stability in degrading AFB1. After treatment in a 100°C boiling water bath for 20 minutes, the degradation rate of 2 μg / mL AFB1 remained stable at over 70% over 48 hours. At 70°C and a pH of 11, 2 μg / mL AFB1 was completely degraded over 48 hours. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 The figure shows the colony morphology of the strain HS-1 provided by the present invention; wherein, (a) shows the colony morphology on a plate; and (b) shows the bacterial morphology under a microscope.

[0045] Figure 2 This is a phylogenetic tree of the strain HS-1 provided by the present invention.

[0046] Figure 3 The present invention provides a standard curve obtained by detecting an AFB1 standard product using HPLC.

[0047] Figure 4 The present invention provides a method for detecting the AFB1 content before and after the reaction of the HPLC test of the supernatant filtrate of the Bacillus paramycoides HS-1 fermentation after high temperature treatment; wherein, (a) is the AFB1 content after 0 hours of reaction; (b) is the AFB1 content after 48 hours of reaction.

[0048] Figure 5 The effects of reaction conditions on the degradation of AFB1 by the supernatant filtrate of Bacillus paramycoides HS-1 fermentation; among them, (a): the effect of temperature; (b): the effect of pH; (c): the effect of metal ions and EDTA. DETAILED DESCRIPTION

[0049] The present invention is described in further detail below with reference to the examples and accompanying drawings, but the present invention is not limited thereto. Experimental methods in the following examples, where specific experimental conditions are not specified, generally followed conventional experimental conditions or those recommended by the manufacturer. Materials and reagents used were commercially available unless otherwise specified.

[0050] Example 1. Screening, separation and purification of strain HS-1

[0051] AFB1-contaminated soil sample from peanut-growing farmland of Guangdong Academy of Agricultural Sciences was sieved through a 2 mm sieve. 1 g of soil was added to 50 mL of LB liquid medium containing 2 μg / mL AFB1 and cultured at 30°C and 180 rpm for 7 days. 5 mL of the culture medium was transferred to fresh LB liquid medium containing 2 μg / mL AFB1 and repeated three times. The culture medium after subculture enrichment was diluted 10-fold (to 10 -6 ), take 10 -3 ~10 -6 The dilution was spread on a 2 μg / mL AFB1-LB agar plate and cultured at 30°C for 72 h. A single colony was picked and streaked three times to obtain a pure culture strain, which was designated as HS-1.

[0052] Example 2: Identification of isolated strains

[0053] (1) Colony morphology identification

[0054] Streak strain HS-1 onto LB plates and incubate at 30°C for 24 hours. Observe and record the characteristics of HS-1 colonies. Separately, select a single HS-1 colony and inoculate it into 20 mL of LB liquid medium. Incubate at 30°C and 180 rpm until the logarithmic growth phase (approximately 12 hours). Smear the culture, flame-fix, and stain with crystal violet solution for 1 minute. Wash with water, then add Gram iodine solution for 1 minute. Wash with water, decolorize with 95% ethanol for 30 seconds, counterstain with safranin solution for 1 minute, rinse, and air-dry before observing cell morphology under an inverted microscope.

[0055] The morphological characteristics of HS-1 provided by the present invention are as follows: after the strain HS-1 is streaked onto an LB agar plate and cultured at 30°C for 24 hours, the colonies are round or with irregular edges, about 3 to 5 mm in diameter, with a dry and rough surface, a typical waxy texture, a grayish white color, a convex center, and thin edges ( Figure 1 (a) in the figure). Gram staining showed that HS-1 bacteria were purple, belonging to Gram-positive bacteria, and the cells were short chain or rod-shaped ( Figure 1 (b) in the figure.

[0056] (2) 16S rDNA gene sequence determination

[0057] The whole genome of the strain culture medium was extracted using a genome extraction kit, and the 16S rDNA of strain HS-1 was amplified using the 16S rDNA universal primers 27F: 5'-AGAGTTTGATCCTGGCTCAG-3' and 1492R: 5'-GGTTACCTTGTTACGACTT-3'. The PCR amplification reaction system was 50 μL, including 0.25 μL Ex Taq DNA polymerase, 5 μL 10× Ex Taq Buffer (MgCl2). 2+ Plus), 4 μL dNTP Mixture, 0.5 μL upstream primer (100 mM), 0.5 μL downstream primer (100 mM), 3.75 μL total DNA, and 36 μL ddH2O. The PCR amplification procedure was as follows: initial denaturation at 95°C (5 min); 30 cycles of 95°C (15 s), 50°C (30 s), and 72°C (1 min); and a final extension at 72°C for 10 min. After amplification, the reaction mixture was separated by 1.5% agarose gel electrophoresis and analyzed on a reflectometer to confirm the presence of a band approximately 1.5 kb. The resulting fragment was then sent to Guangzhou Aiki Biotechnology Co., Ltd. for sequencing.

[0058] Homology searches for the 16S rDNA of strain HS-1 were performed using NCBI BLASTn (https: / / blast.ncbi.nlm.nih.gov) using the "16S ribosomal RNA sequences (Bacteria and Archaea)" database. Model strain 16S rDNA sequences with identical (100%), similar (>99%), or distant (<95%) identities were selected for multiple sequence alignment using MUSCLE 3.8.31 with 100 iterations. Lowly conserved regions were removed using Gblocks 0.91b (parameters: 50% gap tolerance and minimum block length of 5). A phylogenetic tree was constructed using the neighbor-joining method in MEGA 11.0.13. Topological stability was verified with 1000 bootstrap replicates. Tree optimization was performed using FigTree 1.4.4. Nodes were annotated with bootstrap support, and the genetic distance scale was set to 0.005.

[0059] The 16S rDNA of strain HS-1 was amplified by PCR and sequenced to obtain a 1436-bp sequence (SEQ ID NO: 1). Comparison with the GenBank database showed that the sequence was 100% identical to the model strain MCCC 1A04098 / NH24A2 of Bacillus paramycoides (accession number NR_157734.1). Phylogenetic trees were constructed by selecting 16S rDNA sequences of other model strains with the same, similar, or lower identity from the alignment results. Figure 2 The results showed that HS-1 clustered with Bacillus mycoides strains MCCC 1A04098 and JYZ-SD5 into a highly supported branch (bootstrap value 88), forming an independent evolutionary cluster. In contrast, HS-1 was more closely related to Bacillus cereus ATCC 14579 and CCM 2010, and Bacillus thuringiensis IAM 12077 and ATCC 10792 (bootstrap value 62), while it was more genetically distant from Bacillus subtilis 168 and NBRC 13719 (bootstrap value <50%). The results are consistent with the species differentiation law within the genus Bacillus. Bacillus pseudomycoides is highly conserved in phylogeny with the B. cereus group, but its unique 16S rDNA sequence characteristics (such as base differences in the V3 variable region) support its independent species status.

[0060] The present invention finally identified the strain to the genus, and the results showed that the strain was identified as Bacillus paramycoides HS-1, and its homology with Bacillus paramycoides was 100%. In the present invention, the obtained strain was named Bacillus paramycoides HS-1.

[0061] (3) Preservation of bacterial strains

[0062] Prepare the culture into 30% glycerol tubes and store them in a -80°C freezer until use. The above strains were deposited on June 13, 2025, at the Guangdong Provincial Microbiological Culture Collection Center (GDMCC), located at the Institute of Microbiology, Guangdong Academy of Sciences, Building 59, 5th Floor, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Province, with the culture collection number GDMCC No: 66506.

[0063] Example 3: Application of Bacillus paramycoides HS-1 in Degrading AFB1

[0064] (1) HPLC measurement of AFB1 degradation rate

[0065] 500 μL of the culture supernatant was added to 500 μL of chromatography-grade methanol. The mixture was vortexed for 1 minute to thoroughly mix and terminate the reaction. The supernatant was then centrifuged at 12,000 rpm for 5 minutes. The supernatant was filtered through a 0.22 μm nylon 66 filter into a vial. The AFB1 content in the degraded sample was determined by HPLC with a UV detector. High-performance liquid chromatography (HPLC) was used to determine AFB1 using an Agilent ZORBAX Eclipse SB-C18 column (4.6 × 250 mm, 5 μm) with a mobile phase of acetonitrile / water (40 / 60 v / v). The column temperature was 30°C, the flow rate was 1 mL / min, and the detector wavelength was 360 nm. The degradation rate of AFB1 in the sample was calculated using the following formula: degradation rate (%) = (1-A1 / A2) × 100%, where A1 is the residual concentration of AFB1 in the sample (μg / mL) and A2 is the initial concentration of AFB1 in the sample (μg / mL).

[0066] (2) Drawing of AFB1 standard curve

[0067] The AFB1 stock solution with a concentration of 100 μg / mL was diluted with chromatographic grade methanol to obtain AFB1 standard solutions with concentrations of 0.2, 0.5, 1, 2, and 5 μg / mL. The standard solutions were determined according to the detection method described above. Each concentration was determined three times. The AFB1 concentration (μg / mL) was used as the horizontal axis and the AFB1 peak area detected by HPLC was used as the vertical axis. The correlation coefficient was calculated and a standard curve was drawn, as shown in the figure. Figure 3 As shown, the correlation coefficient R 2 =0.9999.

[0068] (3) AFB1 degradation reaction

[0069] Bacillus paramycoides HS-1 was inoculated into LB liquid culture medium containing 2 μg / mL AFB1 and cultured at 30°C and 180 rpm for 16 h. 2 mL of the culture medium was centrifuged at 12,000 rpm for 5 min. 500 μL of the supernatant was taken and the residual AFB1 was detected according to the method described in step (1) and the degradation rate was calculated. The degradation rate was 38.29%.

[0070] Preparation of seed solution: First activate Bacillus paramycoides HS-1, then pick a single colony and add it to NB broth medium, culture at 30°C and 180 rpm for 12 hours to obtain seed solution.

[0071] The seed liquid of Bacillus paramycoides HS-1 was inoculated into NB broth medium at a 5% (v / v) inoculation rate. After culturing at 30°C and 180 rpm for 24 h, the culture medium was centrifuged at 10,000 rpm for 10 min at 4°C. The supernatant was filtered through a 0.22 μm sterile filter membrane and transferred to a sterile centrifuge tube. It was stored at 4°C until use and recorded as the fermentation supernatant filtrate.

[0072] AFB1 was added to the fermentation supernatant filtrate at a final concentration of 2 μg / mL and incubated in a shaker at 30°C and 180 rpm for 48 hours. Simultaneously, the fermentation supernatant filtrate was placed in a 100°C boiling water bath for 20 minutes. After cooling to room temperature, the same concentration of AFB1 was added and incubated under the same conditions for 48 hours. Fermentation supernatant filtrate without AFB1 was used as a negative control, and NB broth containing 2 μg / mL AFB1 was used as a positive control.

[0073] The AFB1 residue was detected and the degradation rate was calculated according to the method described in step (1). The degradation rate of HS-1 fermentation supernatant filtrate for 2 μg / mL AFB1 was 50.47% in 48 h, while the degradation rate of HS-1 fermentation supernatant filtrate for 2 μg / mL AFB1 was 77.17% in 48 h. The HPLC detection charts of AFB1 degradation of the fermentation supernatant filtrate after high temperature treatment for 0 h and 48 h were shown as follows. Figure 4 (a) and Figure 4 Therefore, the degradation activity of HS-1 against AFB1 mainly exists in the extracellular supernatant, and the supernatant filtrate of Bacillus mycoides HS-1 fermentation shows obvious heat activation properties.

[0074] In addition, after high temperature treatment, the HS-1 fermentation supernatant filtrate could degrade 69.42% of 5 μg / mL AFB1 in 48 h; after high temperature treatment, the HS-1 fermentation supernatant filtrate could degrade 60.98% of 10 μg / mL AFB1 in 48 h.

[0075] (4) AFB1 degradation reaction conditions

[0076] In order to evaluate the optimal reaction temperature of the fermentation supernatant filtrate of Bacillus mycoides HS-1, a temperature gradient was set: 10°C, 20°C, 30°C, 40°C, 50°C, 60°C, and 70°C. Specifically, the fermentation supernatant filtrate obtained in step (3) was placed in a 100°C boiling water bath for 20 minutes. After cooling to room temperature, AFB1 was added at a final concentration of 2 μg / mL and placed in a shaker at different temperatures and 180 rpm for 48 hours. The residue of AFB1 was detected and the degradation rate was calculated. The degradation rates were 59.31%, 65.51%, 76.12%, 80.97%, 89.25%, 97.04%, and 99.02%, respectively. Figure 5 (a) in Figure 2). It can be seen that the degradation rate is highest at 70°C.

[0077] In order to evaluate the optimal reaction pH of the fermentation supernatant filtrate of Bacillus mycoides HS-1, the fermentation supernatant filtrate obtained in step (3) was placed in a 100°C boiling water bath for 20 minutes. After cooling to room temperature, the pH gradient was adjusted to 3, 4, 5, 6, 7, 8, 9, 10, and 11. Specifically, AFB1 was added to the fermentation supernatant filtrate at different pH values ​​at a final concentration of 2 μg / mL and placed in a shaker at 30°C and 180 rpm for 48 hours. The residue of AFB1 was detected and the degradation rate was calculated. The degradation rates were 13.72%, 15.22%, 27.39%, 37.29%, 68.86%, 84.53%, 87.29%, 89.66%, and 96.59%, respectively. Figure 5 (b) in Figure 2). It can be seen that the degradation rate is highest when the pH is 11.

[0078] In summary, the fermentation supernatant filtrate obtained in step (3) was placed in a 100°C boiling water bath for 20 minutes. After cooling to room temperature, the pH was adjusted to 11, and AFB1 was added to a final concentration of 2 μg / mL. The mixture was then shaken at 70°C and 180 rpm for 48 hours. The residual AFB1 was detected and the degradation rate was calculated. The results showed that 2 μg / mL AFB1 could be completely degraded at 70°C, pH 11, and 48 hours.

[0079] The factors affecting the AFB1 degradation activity of the fermentation supernatant filtrate of Bacillus mycoides HS-1 were also evaluated. The fermentation supernatant filtrate obtained in step (3) was placed in a 100°C boiling water bath for 20 minutes. After cooling to room temperature, LiCl, CuCl2, CaCl2, MnSO4, MgCl2, ZnCl2, FeCl3 and EDTA were added at a final concentration of 100mM, and AFB1 was added at a concentration of 2μg / mL. The mixture was placed in a shaker at 30°C and 180rpm for 48 hours. The treatment without adding metal ions or EDTA was used as the control (Control). The residue of AFB1 was detected and the degradation rate was calculated. The degradation rates were 68.56%, 77.73%, 44.38%, 54.22%, 61.46%, 0.54%, 71.86%, 4.92% ( Figure 5 Compared with the control (74.34%), the AFB1 degradation activity of the HS-1 fermentation supernatant filtrate could be significantly increased by Cu 2+ Improved by EDTA and Zn 2+ , Ca 2+ 、Mn 2+ Mg 2+ Significant inhibition.

[0080] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be considered as equivalent replacement methods and are included in the scope of protection of the present invention.

Claims

1. A strain of aflatoxin B1-degrading bacteria, characterized by: The name is Bacillus paramycoides HS-1, and it was deposited on June 13, 2025 in the Guangdong Microbial Culture Collection Center, Institute of Microbiology, Guangdong Academy of Sciences, 5th Floor, Building 59, Compound 100, Xianlie Middle Road, Guangzhou, with the deposit number: GDMCC No: 66506.

2. An aflatoxin B1 degradation reagent, characterized in that: The degradation agent comprises at least one of the Bacillus mycoides HS-1, Bacillus mycoides HS-1 fermentation broth, Bacillus mycoides HS-1 fermentation broth supernatant and Bacillus mycoides HS-1 fermentation supernatant filtrate as claimed in claim 1.

3. The aflatoxin B1 degradation reagent according to claim 2, characterized in that: The Bacillus mycoides HS-1 fermentation liquid is prepared by liquid fermentation of the Bacillus mycoides HS-1 described in claim 1.

4. The aflatoxin B1 degradation reagent according to claim 3, characterized in that: The method is to inoculate the seed liquid of Bacillus mycoides HS-1 into NB broth culture medium for fermentation to obtain fermentation liquid.

5. The aflatoxin B1 degradation reagent according to claim 4, characterized in that: The inoculation amount of the seed solution is 5-10%; The fermentation culture conditions are 28-30° C., 180-200 r / min, and 24-72 hours.

6. The aflatoxin B1 degradation reagent according to any one of claims 2 to 5, characterized in that: The Bacillus mycoides HS-1 fermentation broth supernatant is prepared by centrifuging the Bacillus mycoides HS-1 fermentation broth; The supernatant filtrate of the Bacillus mycoides HS-1 fermentation broth is prepared by filtering the supernatant of the Bacillus mycoides HS-1 fermentation broth.

7. The aflatoxin B1 degradation reagent according to claim 6, characterized in that: The centrifugal conditions are 2-8°C, 8000-10000 rpm, and 8-10 min; The filtration is performed using a 0.22 μm sterile filter membrane.

8. Use of the aflatoxin B1-degrading bacteria according to claim 1 or the aflatoxin B1-degrading reagent according to any one of claims 2 to 7 for non-disease treatment purposes in degrading aflatoxin B1.

9. The use according to claim 8, characterized in that: The steps include: (1) inoculating the Bacillus mycoides HS-1 described in claim 1 into LB liquid culture medium containing AFB1 and culturing; or, (2) inoculating the seed liquid of the Bacillus mycoides HS-1 described in claim 1 into NB broth medium, and after fermentation, centrifuging the culture liquid, collecting the fermentation liquid supernatant, and filtering the fermentation liquid supernatant to obtain a fermentation supernatant filtrate; (3) adding AFB1 to the fermentation supernatant filtrate of step (1) and reacting; (4) The fermentation supernatant filtrate of step (1) was heated, cooled to room temperature, and then AFB1 was added and reacted.

10. The use according to claim 9, characterized in that: In step (1), the culture conditions are 28-30° C., 180-200 r / min, and 16-48 h; In step (2), the inoculation amount of the seed solution is 5-10%; In step (2), the fermentation culture conditions are 28-30° C., 180-200 r / min, and 24-72 h; In step (2), the centrifugation conditions are 2-8°C, 8000-10000 rpm, and 8-10 min; In step (2), the filtration is performed using a 0.22 μm sterile filter membrane; In steps (3) and (4), the reaction conditions are 28-30°C, 180-200 r / min, and 48-72 h; In step (4), the heating treatment condition is 95-100° C. for 15-20 min.

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