Bacillus pumilus as well as preparation method and application thereof

Abstract

The invention relates to the technical field of microbial screening and biodegradation, and discloses bacillus pumilus as well as a preparation method and application thereof. Microorganisms are enriched from vinasse compost, the bacillus pumilus DF-1 is obtained through TOC degradation rate screening, the bacillus pumilus DF-1 is preserved in Guangdong Microbial Culture Collection Center on September 22, 2025, and the preservation number is GDMCC NO: 67011. The strain efficiently degrades organic matters of the vinasse under the acidic conditions that the temperature is 50-55 DEG C and the pH value is smaller than or equal to 5, lime does not need to be added for neutralization, the problems of nutrient loss and mineral imbalance existing in a traditional method are solved, and an efficient microbial technical scheme is provided for vinasse resourceful treatment.

Description

Technical Field

[0001] This invention relates to the field of microbial screening and biodegradation technology, specifically to a short-lived Bacillus strain, its preparation method, and its applications. Background Technology

[0002] The production of baijiu (Chinese liquor) generates a large amount of lees waste (pH < 5) annually, which is highly acidic, high in lactic acid, and rich in recalcitrant organic matter. Traditional composting techniques suffer from insufficient acid resistance and high-temperature adaptability of microbial agents, resulting in a long composting period of 30-60 days. Existing high-temperature microbial agents are mostly designed for conventional organic matter and have poor adaptability to the unique acidic environment (pH = 3-5) and high-temperature conditions (50-55℃) of lees. Studies have shown that Bacillus licheniformis and Rhizopus microphylla can maintain intracellular pH homeostasis, and Aspergillus niger secretes urease to neutralize the acidic environment. However, there is limited research on the degradation capabilities of these strains at temperatures above 50℃, and there is a lack of dedicated strains for efficiently degrading organic matter in lees. Current techniques often add lime to neutralize acidity, but this leads to nitrogen loss, decomposition of heat-sensitive substances, and an imbalance in the calcium-phosphorus ratio, reducing the nutritional value of the compost. Furthermore, uneven lime dispersion increases costs. Summary of the Invention

[0003] This invention aims to solve the technical problems of existing composting microbial agents, such as low degradation efficiency in high-temperature acidic environments above 50℃, reliance on lime neutralization leading to decreased compost quality, and long composting cycles. It provides a strain that can stably grow and efficiently degrade organic matter in high-temperature environments above 50℃ and acidic environments with a pH of 3-5; avoids nutrient loss and operational costs associated with lime neutralization; shortens the composting cycle of distiller's grains; and improves the conversion efficiency of organic matter.

[0004] To achieve the above objectives, the first aspect of this application provides a *Bacillus pumilus*, wherein the *Bacillus pumilus* (… Bacillus licheniformis DF-1 was deposited at the Guangdong Microbial Culture Collection Center on September 22, 2025, with accession number GDMCC NO:67011. Furthermore, the *Bacillus pumilus* grows in a temperature range of 50-55°C and / or an acidic environment with pH ≤ 5, and degrades the organic matter in the distiller's grains.

[0005] Secondly, this application also provides a method for preparing the aforementioned Bacillus pumilus, comprising the following steps: S1. Enrichment culture: Inoculate compost samples and distiller's grains samples into bacterial enrichment culture medium and culture at 50℃ and 160 r / min for 5-7 days with shaking. S2. Separation and purification: Take the enriched bacterial solution and inoculate it onto the bacterial solid culture medium using the dilution and spread method. Incubate at 50℃ for 2-3 days and pick out a single colony. S3. Expansion and preservation: Single colonies were inoculated into LB liquid medium for expansion, and after subculturing, they were preserved in glycerol at -80℃.

[0006] Furthermore, the bacterial enrichment culture medium contains 5.0 g / L peptone, 5.0 g / L yeast extract, and 3.0 g / L NaCl.

[0007] Furthermore, the bacterial solid culture medium contains 5.0 g / L peptone, 5.0 g / L yeast extract, 3.0 g / L NaCl, and 15-20 g / L agar.

[0008] Thirdly, this application also provides a composting high-temperature degradation microbial agent, comprising the Bacillus fabricus DF-1 described in this invention, suitable for the organic matter degradation and maturation promotion of distiller's grains compost. Fourthly, the application of Bacillus pumilus DF-1 in the degradation of organic matter in distiller's grains compost.

[0009] Fifthly, the method of degrading organic matter in distiller's grains using the aforementioned Bacillus DF-1 involves inoculating the strain at a rate of 2% to 3% of the total volume of the composting system and promoting TOC degradation at 50 to 55°C.

[0010] Furthermore, an acidity correction experiment was conducted, in which the pH of the culture medium was periodically adjusted to the initial acidity value of 3.6 using 0.1 mol / L dilute hydrochloric acid during the culture process.

[0011] Furthermore, the culture temperature is 55°C.

[0012] The isolated and purified strain DF-1 was subjected to 16S rRNA sequencing. The obtained gene sequence was analyzed using BLAST in NCBI. The gene sequence of strain DF-1 was compared with that of Bacillus pumilus (…). Bacillus licheniformis The similarity between the strain and the target strain is over 98%, therefore the strain identified is *Bacillus pumilus* (B. pumilus). Bacillus licheniformis ).

[0013] Compared with existing technologies, the composting thermophilic degrading bacteria and their applications provided by this invention have the following significant beneficial effects: 1. Excellent high-temperature tolerance and efficient adaptability to acidic environments: The short-spore Bacillus strains obtained by screening in this invention ( Bacillus licheniformis DF-1 can grow well under high temperature conditions of 50℃ to 55℃. It maintains high metabolic activity and can tolerate and actively regulate the unique acidic environment of the distiller's grains (initial pH 3.6) to raise it to the neutral range (pH 7.2-7.6), effectively overcoming the technical bottleneck of traditional microbial agents whose activity is inhibited under high temperature and acidic conditions.

[0014] 2. Significantly improves organic matter degradation efficiency and greatly shortens the composting cycle: After applying this strain, the total organic carbon (TOC) degradation rate in the distiller's grains composting system can reach up to 71.97%, and the degradation is mainly concentrated in the first 48 hours. Compared with the 30-60 day maturation cycle of traditional composting technology, this invention can greatly shorten the composting time and improve the processing efficiency.

[0015] 3. Avoids the negative effects of using lime for neutralization: This strain has the ability to regulate pH, reducing or avoiding the traditional operation of adding lime to adjust acidity. This prevents problems such as nitrogen loss, decomposition of heat-sensitive nutrients (such as B vitamins and amino acids), and imbalance of calcium and phosphorus ratio caused by adding lime, and helps to preserve and improve the fertilizer value and quality of compost products.

[0016] 4. Providing a specialized and efficient technical solution for the resource utilization of distiller's grains: The strain of this invention originates from the distiller's grains composting environment and is a functional strain specifically screened for the characteristics of distiller's grains waste. It has a specific ability to degrade refractory organic matter such as lignocellulose in distiller's grains, providing an efficient and specialized microbial solution for the high-value resource utilization of distiller's grains, with significant environmental and economic benefits.

[0017] 5. Stable strain performance and simple application: This strain can be propagated through conventional culture media and can be stably preserved for a long time through glycerol preservation. The production process is simple and low-cost, making it easy to promote and apply on a large scale in actual composting projects. Attached Figure Description

[0018] Figure 1 The graph shows the change in TOC degradation rate of the composting bacteria enrichment solution. Figure 2 The pH adaptability of Bacillus pumilus to the primary screening medium at 50 ℃ was determined. Figure 3 The growth curve of Bacillus pumilus in the primary screening medium at 50 ℃; Figure 4 The change in TOC content of the initial screening culture medium at 50 ℃; Figure 5 The pH adaptability of Bacillus pumilus to the primary screening medium at 55 ℃ was determined. Figure 6 The growth curve of Bacillus pumilus in the primary screening medium at 55 ℃; Figure 7 The change in TOC content of the initial screening culture medium at 55 ℃; Figure 8 The growth curves of Bacillus pumilus in the primary screening medium under periodic acid regulation are shown. Figure 9The changes in TOC content in the primary screening medium under periodic acidic regulation; Figure 10 This graph shows the change in TOC degradation rate of Bacillus pumilus in the primary screening medium. Figure 11 The TOC degradation rate of Bacillus pumilus in the primary screening medium. Detailed Implementation

[0019] The following provides specific operating steps and detailed explanations for the above examples, but these do not constitute a limitation on the technical solution of the present invention.

[0020] Example 1: Enrichment of functional bacteria To screen out well-growing thermophilic bacteria for composting, it is necessary to first conduct bacterial enrichment culture to increase the number of strains, which facilitates the isolation and purification of microorganisms.

[0021] The enrichment sample consisted of 5 g of compost product, 1 g of distiller's grains, and 150 mL of bacterial enrichment medium. The bacterial enrichment medium was formulated as follows: 5.0 g peptone, 5.0 g yeast extract, 3.0 g NaCl, 1000 mL deionized water, natural pH, sterilized at 121℃ for 20 min.

[0022] Two control groups and five parallel experimental groups were set up for bacterial enrichment culture. 250 mL Erlenmeyer flasks sterilized by autoclaving at 121 °C for 20 min were used to hold the culture medium. The compost, distiller's grains, and bacterial enrichment medium were mixed thoroughly and incubated in a 50 °C shaking incubator for 5–7 days at a shaking speed of 160 rpm. -1 There was no light.

[0023] Take 5 mL of bacterial culture every 24 hours and use 5000 r min. -1 Centrifuge for 10 min, collect the supernatant and determine the TOC content at that time using a total organic carbon analyzer, and calculate the TOC degradation rate. Figure 1 It can be seen that the TOC degradation rate of the bacterial enrichment solution decreased slightly in the first two days of enrichment, but from the second day to the fifth day, it showed an upward trend, with the highest TOC degradation rate reaching 81.60%. The TOC degradation rate gradually stabilized from the fifth day onwards. Therefore, the second group of bacterial enrichment solutions (final degradation rate of 79.44%) was selected for separation and purification.

[0024] Example 2: Isolation, purification, and preservation of bacterial strains Take the bacterial enrichment solution from Example 1, pipette 1 mL into a 10 mL centrifuge tube, add sterile deionized water sterilized by autoclaving at 121°C for 20 min, and shake thoroughly to obtain a diluted bacterial solution with a concentration of 10⁻¹.-1 Take 1 mL of the diluted bacterial solution and add it to another 10 mL centrifuge tube containing 9 mL of sterile deionized water to obtain 10 mL of the diluted solution. -2 Diluted bacterial solutions of varying concentrations were prepared. This process was repeated to create dilutions with concentrations of 10-10. -1 10 -2 , ... , 10 -9 Diluted bacterial solution of a certain concentration.

[0025] The prepared diluted bacterial suspension (200 μL) was inoculated onto the surface of a bacterial solid culture medium using the dilution plating method. A glass spreader, sterilized by flaming with an alcohol lamp after being soaked in 75% ethanol, was then evenly spread onto the solid culture medium. All spread media were then inverted and placed in a 50 °C incubator for 2–3 days.

[0026] The formula for bacterial solid culture medium is as follows: 5.0 g peptone, 5.0 g yeast extract, 3.0 g NaCl, 15-20 g agar, 1000 mL deionized water, natural pH, sterilized at 121 ℃ for 20 min.

[0027] Select healthy single bacterial strains and place them in 30 mL of bacterial LB medium. Shake well and incubate at 50 °C for 160 rpm. -1 The bacterial culture was expanded in a shaking incubator for 2-3 days. Then, 200 μL of the expanded bacterial culture was spread onto the surface of a bacterial solid culture medium and incubated upside down in a 50 °C constant temperature incubator for 2-3 days. To obtain well-grown single bacteria, the above operation can be repeated multiple times.

[0028] The formulation of the bacterial LB medium involved is as follows: 10.0 g tryptone, 5.0 g yeast extract, 10.0 g NaCl, 1000 mL deionized water, natural pH, sterilized at 121 ℃ for 20 min.

[0029] Select healthy bacterial cells and inoculate them into 30 mL of bacterial LB medium. Incubate at 50 °C and 160 rpm. -1 Subculture them in a shaking incubator for 3-4 days.

[0030] After subculturing, the bacterial culture was incubated at 5000 rpm. -1 Centrifuge for 10 min, discard the supernatant, and retain 1 mL of bacterial suspension. Use a pipette to draw 800 μL of the resuspended bacterial suspension and add it to 800 μL of 80% glycerol that has been autoclaved at 121 °C for 20 min. Mix thoroughly and store at -80 °C for long-term storage.

[0031] Example 3: Identification of Functional Bacteria The colony characteristics of strain DF-1 are as follows: Under high temperature conditions of 50 ℃, the colonies on the surface of the bacterial solid culture medium grow well. The colonies are elliptical to columnar, located in the center of the cell or slightly off-center. After spore formation, the cell does not swell. The surface is rough and opaque, dirty white or slightly yellow.

[0032] After PCR amplification, the complete 16S rRNA sequence of DF-1 bacteria was obtained as shown in SEQ ID NO. 1: After comparison, the gene sequence is identical to that of Bacillus pumilus (Bacillus subtilis). Bacillus licheniformis The strain showed a similarity of over 98%, therefore it was identified as Bacillus pumilus.

[0033] Example 4: Application of functional bacteria in the degradation of organic matter in distiller's grains 0.1 mL of the bacterial culture medium from Example 3 was pipetted into 10 mL of bacterial LB medium and incubated at 50 °C and 160 r / min. -1 The cultures were incubated in a vibrating incubator for 2-3 days. Three replicates were prepared for each bacterial strain, and a control group was also included. After incubation, the cultures were rotated at 5000 r·min. -1 Centrifuge the bacterial culture for 10 min, remove the supernatant, add sterile deionized water, and continue until the color of the culture is visible on a UV-Vis spectrophotometer. OD 600 The value is around 1.0, thus obtaining a bacterial suspension. The bacterial suspension is added to 100 mL of primary screening medium at an inoculation rate of 2%–3%.

[0034] The formula for the initial screening culture medium is as follows: 25 g of distiller's grains sample, 5.0 g of NaCl, 1000 mL of deionized water, natural pH, sterilized at 121℃ for 30 min.

[0035] Take 5 mL of culture medium every 8 hours for use OD 600 Determination of organic matter and degradation of the culture medium. The culture medium was measured using a UV-Vis spectrophotometer. OD 600 The pH value of the culture medium was measured using a portable multi-parameter water quality analyzer. The bacterial culture was then heated at 5000 rpm. -1 Centrifuge for 10 min, and collect the supernatant for TOC determination using a total organic carbon analyzer. Finally, calculate the TOC degradation rate and degradation percentage at each time point in the initial screening medium. The degradation of organic matter in distiller's grains compost by thermophilic bacteria is investigated based on the TOC degradation rate and degradation percentage.

[0036] Depend on Figure 2The *Bacillus pumilus* strain can adjust the initial pH of the screening medium from 3.6 to approximately 7.2, with significant adjustment within the first 32 hours. Afterward, the pH of the medium tends to stabilize, indicating that the strain of this invention can adapt to the high-temperature, acidic environment of the screening medium and possesses the ability to regulate pH. Within the first 16 hours, *Bacillus pumilus* rapidly proliferates and grows (…). Figure 3 At this point, the growth slows down, and from 48 hours onwards, the number of strains gradually decreases. Therefore, this strain can reproduce and grow in a high-temperature, acidic environment. Furthermore, *Bacillus pumilus* can significantly degrade TOC content, with a degradation rate reaching up to 71.7%. Figure 4 and Figure 10 Specifically, in the first 32 hours, the TOC content in the initial screening medium decreased significantly, and then tended to stabilize. Within the first 8 hours, the TOC degradation rate increased rapidly, then gradually slowed down after 8 hours. Figure 11 ).

[0037] Example 5: Application of functional bacteria in the degradation of organic matter in distiller's grains at 55 °C The culture conditions of Example 4 were changed to 55 °C, while the other methods for preparing the bacterial suspension, culture conditions, and assays remained unchanged. This was to further explore the high-temperature tolerance and organic matter degradation ability of the strain of this invention above 50 °C.

[0038] In a culture system at 55 °C, Bacillus pumilus can adjust the pH of the initial screening medium from 3.6 to 7.6, with the fastest adjustment occurring within the first 32 hours, after which the regulatory ability enters a stable phase. Figure 5 The number of Bacillus subtilis increases rapidly in the first 16 hours, then enters a stable growth phase, and begins to decrease after 48 hours. Figure 6 Bacillus pumilus efficiently degrades the TOC in the primary screening medium, and the TOC content stabilizes after 48 h. Figure 7 The degradation rate can reach up to 71.97%. Figure 10 Bacillus pumilus exhibits the fastest degradation rate of organic matter in distiller's grains during the first 8 hours, followed by a decrease in degradation rate but a steady degradation process. Figure 11 ).

[0039] Example 6: Application of functional bacteria in the degradation of organic matter in distiller's grains at 55 °C with acidity correction The culture conditions of Example 4 were changed to 55 °C, and the pH of the initial screening medium was adjusted to approximately 3.6 with 0.1 mol / L dilute hydrochloric acid after sampling every 8 hours. The other methods for preparing the bacterial suspension, culture conditions, and assays remained unchanged. This allows for further research on the adaptability of Bacillus pumilus to acidic environments and its ability to degrade organic matter, while also mitigating the drawbacks of lime-based acidification in composting.

[0040] from Figure 8 It was observed that *Bacillus pumilus* multiplied rapidly in the first 24 hours, then its population growth plateaued, and the number of strains began to decrease after 48 hours. Furthermore, the TOC content of the initial screening medium degraded rapidly and significantly, also stabilizing after 48 hours, with the highest degradation rate reaching 70.03%. Figure 9 and Figure 10 Its degradation rate increases rapidly within the first 8 hours, and then the degradation rate fluctuates steadily thereafter. Figure 11 ).

[0041] In summary, the selected Bacillus pumilus (Bt) Bacillus licheniformis DF-1 can tolerate high-temperature acidic environments above 50℃ (50℃ and 55℃), and can regulate the pH value from acidic to neutral, and can reproduce and grow rapidly. Moreover, it can efficiently degrade the organic matter content of distiller's grains compost, with a degradation rate as high as 71.7% at 50℃; a degradation rate as high as 71.97% at 55℃; and a degradation rate as high as 70.03% at 55℃ under acidic conditions. This microorganism is *Bacillus pumilus* (…). Bacillus licheniformis DF-1 can be applied to the composting process of distiller's grains to solve the technical defects of lime acidity adjustment and improve the resource utilization of distiller's grains.

[0042] Application Experiment: To clarify the advantages of the microbial agent of this invention in actual fertilizer production, the applicant cooperated with a local organic fertilizer manufacturer to conduct a comparative experiment. With "simulated factory composting conditions" as the core, the actual application effects of the two groups of microbial agents were compared by controlling variables. The method is highly operable and meets production needs.

[0043] I. Experimental Materials and Equipment 1. Experimental materials Experimental bacterial agent: The Bacillus pumilus DF-1 bacterial agent described in this application was expanded and cultured according to the preparation method of claim 3, and the OD600 value of the bacterial suspension was adjusted to 1.0.

[0044] Control group microbial agent: The factory currently uses the commonly used composting fermentation microbial agent (organic fertilizer fermentation agent purchased from Shandong Beijia Biotechnology Co., Ltd.), and uses it according to its recommended method.

[0045] Composting materials: fresh distiller's grains (pH 3.5-4.5, record the initial value after TOC content determination), straw (crushed to 2-3cm, adjust the carbon-nitrogen ratio to 25:1), the amount of raw materials should be prepared according to the actual production ratio of the factory.

[0046] 2. Experimental equipment Factory standard composting fermentation chambers (2 units, identical in size, with real-time temperature and pH monitoring functions).

[0047] Total organic carbon (TOC) analyzer, portable pH meter, UV-Vis spectrophotometer.

[0048] Sampling tools (sterile sampling bags, sampling spoons), electronic balance.

[0049] II. Experimental Methods 1. Composting system setup (2 parallel groups, 3 replicates per group) Experimental group: Take 1000kg of distiller's grains and 200kg of crushed straw, add DF-1 bacterial suspension at an inoculation rate of 2% of the total volume of the composting system, mix evenly, and then transfer to the fermentation chamber.

[0050] Control group: Take 1000kg of distiller's grains and 200kg of crushed straw with the same ratio, add the commonly used microbial agent suspension in the factory at the same inoculation amount (2%), mix evenly and then transfer to another fermentation chamber.

[0051] Both fermentation chambers were equipped with the same ventilation conditions (ventilation volume 0.5m³). 3 / (m 3 •h)) and turning frequency (turning the pile once every 24 hours), simulating the actual production parameters of the factory.

[0052] 2. Sampling and testing (cycle: 0-15 days, 1 sample every 24 hours) Temperature monitoring: Record the highest daily temperature in the two fermentation chambers and observe the duration of the high-temperature period (≥50℃).

[0053] pH measurement: Take a compost sample (50g), add deionized water at a ratio of 1:10 and soak for 30 minutes. Measure the pH of the supernatant using a portable pH meter.

[0054] TOC degradation rate determination: Take 10g of compost sample, freeze-dry and grind it, and measure the TOC content using a TOC analyzer. Calculate the degradation rate according to the formula: (initial TOC content - daily TOC content) / initial TOC content × 100%.

[0055] Determining compost maturity: On day 15, the germination index (GI) of the two compost groups was tested. A GI ≥ 80% was considered mature. The time it took for the two groups to reach maturity was recorded.

[0056] 3. Nutrient composition testing (after composting is completed, i.e., when both groups have reached full decomposition). The total nitrogen (Kjeldahl method) and total phosphorus (molybdenum-antimony colorimetric method) contents of the two groups of compost products were determined, and the nutrient retention effects of the two groups of microbial agents were compared under the condition of lime neutralization (traditional method) absence.

[0057] III. Experimental Results Data (I) Basic environmental and degradation efficiency indicators

[0058] (II) Nutritional Indicators of Compost Products

[0059] IV. Data Analysis 1. pH adjustment ability: The experimental group adjusted the initial pH of 3.8 to neutral (7.2) on the 7th day and stabilized at 7.5 on the 10th day without the need to add lime; the control group had a pH of only 5.8 on the 10th day and required additional neutralization.

[0060] 2. Organic matter degradation efficiency: The TOC degradation rate of the experimental group was 71.9% on day 10, which was 47.9% higher than that of the control group (48.6%). The composting cycle was shortened from 20 days to 11 days, and the efficiency was increased by 45%, which has a significant advantage in shortening the composting cycle.

[0061] 3. High temperature adaptability and composting quality: The experimental group started the high temperature period 17 hours earlier and lasted for 5 days longer, showing good tolerance to 50~55℃; after composting, the total nitrogen and total phosphorus contents increased by 39.6% and 29.7% respectively compared with the control group, avoiding nutrient loss caused by lime, with a higher germination index and better product safety.

[0062] 4. Production applicability: The experimental group does not require lime, reducing cost and quality fluctuations; the shortened cycle helps to increase the factory's annual processing capacity.

[0063] V. Conclusion The experimental data above show that the Bacillus pumilus DF-1 inoculant described in this application is significantly superior to commonly used composting inoculants in factories in terms of pH adjustment, organic matter degradation, high temperature adaptation, and nutrient retention in distiller's grains compost, and has the feasibility and superiority for large-scale application in fertilizer production plants.

[0064] The above descriptions are merely embodiments of the present invention, and common knowledge such as specific technical solutions and / or characteristics are not described in detail here. It should be noted that those skilled in the art can make various modifications and improvements without departing from the technical solutions of the present invention, and these should also be considered within the scope of protection of the present invention. These modifications and improvements will not affect the effectiveness of the implementation of the present invention or the practicality of the patent. The scope of protection claimed in this application should be determined by the content of its claims, and the specific embodiments described in the specification can be used to interpret the content of the claims.

Claims

1. A short-lived Bacillus species, characterized in that: The short-spore-forming bacteria ( Bacillus licheniformis DF-1 was deposited at the Guangdong Microbial Culture Collection Center on September 22, 2025, with accession number GDMCC NO:67011.

2. The *Bacillus pumilus* according to claim 1, characterized in that: The *Bacillus pumilus* grows in a temperature range of 50-55°C and / or an acidic environment with pH ≤ 5, and degrades organic matter in the distiller's grains.

3. The method for preparing Bacillus pumilus according to claim 1, characterized in that: Includes the following steps: S1. Enrichment culture: Inoculate compost samples and distiller's grains samples into bacterial enrichment culture medium and culture at 50℃ and 160 r / min for 5-7 days with shaking. S2. Separation and purification: Take the enriched bacterial solution and inoculate it onto the bacterial solid culture medium using the dilution and spread method. Incubate at 50℃ for 2-3 days and pick out a single colony. S3. Expansion and preservation: Single colonies were inoculated into LB liquid medium for expansion, and after subculturing, they were preserved in glycerol at -80℃.

4. The method according to claim 3, characterized in that: The bacterial enrichment medium contains 5.0 g / L peptone, 5.0 g / L yeast extract, and 3.0 g / L NaCl.

5. The method according to claim 3, characterized in that: The bacterial solid culture medium contains 5.0 g / L peptone, 5.0 g / L yeast extract, 3.0 g / L NaCl, and 15-20 g / L agar.

6. A composting high-temperature degradation microbial agent, characterized in that: Containing the Bacillus DF-1 as described in claim 1, it is suitable for promoting the organic matter degradation and composting of distiller's grains.

7. The application of the Bacillus DF-1 of claim 1 in the degradation of organic matter in distiller's grains compost.

8. A method for degrading organic matter in distiller's grains using Bacillus fabricus DF-1 as described in claim 1, characterized in that: The strain was inoculated at 2% to 3% of the total volume of the composting system, and TOC degradation was promoted at 50 to 55°C.

9. The method according to claim 7, characterized in that: In a simulated acidity correction experiment, the pH of the culture medium was periodically adjusted to the initial acidity value of 3.6 using 0.1 mol / L dilute hydrochloric acid during the culture process.

10. The method according to claim 8, characterized in that: The culture temperature is 55℃.

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