Agrobacterium radiobacter and application thereof in ethyl acetate degradation

Abstract

The application discloses a strain of Agrobacterium pleuropneumoniae and application thereof in ethyl acetate degradation, and belongs to the technical field of microorganisms. Agrobacterium pusense The preservation number of the Agrobacterium pleuropneumoniae (F21) is CCTCC NO: M 2026373, the strain is separated from a deodorization treatment system, and the strain is first found to have the function of efficiently degrading ethyl acetate, has fast growth speed, high viable cell count and outstanding ethyl acetate removal capacity. Meanwhile, the strain has good extreme environment tolerance capacity and can normally grow in normal or extreme environments. Therefore, the strain has both efficient degradation capacity and wide environmental adaptability, and has significant industrial application potential.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, and more specifically, to a strain of *Pusamobacillus* and its application in the degradation of ethyl acetate. Background Technology

[0002] Ethyl acetate, also known as ethyl acetate, is an organic compound with the chemical formula C4H8O2. It is an ester with the functional group -COOR (a double bond between carbon and oxygen). It can undergo common ester reactions such as alcoholysis, ammonolysis, transesterification, and reduction. It is mainly used as a solvent, food flavoring, and cleaning and degreasing agent.

[0003] Ethyl acetate, a widely used organic solvent, poses environmental hazards throughout its production, use, and disposal. Atmospherically, as a typical volatile organic compound (VOC), it reacts with nitrogen oxides under sunlight to generate photochemical smog, and its greenhouse effect is also significant, earning it a place on the list of key controlled pollutants. In water bodies, ethyl acetate is readily degraded by microorganisms, rapidly consuming dissolved oxygen and leading to oxygen depletion, creating "dead water zones" where fish and other aquatic life cannot survive, and exhibiting direct toxicity to aquatic organisms. Its harm to soil and groundwater is even more insidious; once leaked, it rapidly infiltrates and forms a stable pollution plume, migrating long distances with groundwater, making remediation extremely difficult and damaging soil ecosystems, becoming a long-term source of release. Ultimately, these pollutants pose health risks to the human central nervous system and respiratory system through ecosystems and inhalation.

[0004] Traditional treatment methods, such as regenerative thermal oxidizers, suffer from high investment costs and energy consumption. In contrast, biodegradation of VOCs and other harmful substances offers advantages such as high cost-effectiveness, minimal residues or byproducts, and environmental friendliness. Therefore, developing a cost-effective biological treatment technology that can withstand complex conditions such as high concentrations and strong acids / alkalis is an urgent need to address these environmental and industrial challenges.

[0005] In view of this, the present invention is proposed. Summary of the Invention

[0006] The purpose of this invention is to provide a strain of Agrobacterium pusanum and its application in the degradation of ethyl acetate. This strain has both broad-spectrum environmental adaptability and high-efficiency degradation ability, and can stably remove ethyl acetate under extreme conditions such as strong acid, strong alkali and high concentration.

[0007] This invention is implemented as follows: In a first aspect, the present invention provides a strain of *Pseudomonas aeruginosa* (… Agrobacterium pusense ).

[0008] The *Agrobacterium praosporum* of this invention was isolated from a biological deodorization system in Hangzhou, and its taxonomic name is *Agrobacterium praosporum* (…). Agrobacterium pusense F21, its preservation information is as follows: Depository Institution: China Center for Type Culture Collection (CCTCC), Deposit Date: March 4, 2026, Deposit Address: Wuhan University, Wuhan, China, Deposit Number: CCTCC NO: M 2026373.

[0009] Psammobacterium ( Agrobacterium pusense F21 is a Gram-negative bacterium. Colonies grown on LB agar are round, yellow on the front, smooth, with neat and clear edges, easily picked up, and have a short rod-like microstructure. Currently... Agrobacterium The publicly available information regarding pollutant degradation includes the ability to degrade pollutants such as ethyl carbamate and 2,4-di-tert-butylphenol. This invention is the first to discover that the strain of *Psammonium pusanum* has a good ability to remove ethyl acetate.

[0010] Secondly, the present invention provides a microbial inoculant containing the aforementioned *Pusarobacter* (…). Agrobacterium pusense )F21.

[0011] Furthermore, the aforementioned microbial agents, classified by physical form, can be liquid formulations, semi-solid formulations, or solid formulations. Specifically, the types of liquid formulations include, but are not limited to, *Plasmobium* (…). Agrobacterium pusense F21 fermentation broth, concentrated bacterial suspension, bacterial cell suspension, semi-solid preparations including but not limited to bacterial mud and bacterial paste; solid preparations including but not limited to bacterial powder, freeze-dried powder, spray-dried powder, granules, tablets, etc.

[0012] In some embodiments, the microbial agent is a composition, such as *Pusarobacter* (…). Agrobacterium pusense F21 is the main active ingredient of this microbial agent. In addition to the main active ingredient, the microbial agent of this invention may also include excipients or carriers conventional in the art. Excipients may be protective agents, stabilizers, penetrants, or pH adjusters, and carriers may be organic polymers, inorganic materials, or complexes. Those skilled in the art can adjust the dosage form of the microbial agent, as well as the type or amount of excipients and carriers, according to the application scenario and needs. All microbial agents containing *Pseudomonas aeruginosa* (F21) of this invention... Agrobacterium pusense Microbial agents designed with F21 as the main active ingredient are all within the scope of protection of this invention.

[0013] In some embodiments, the method for preparing the above-mentioned microbial agent includes: culturing the above-mentioned *Pusarobacter* (…). Agrobacterium pusense )F21.

[0014] Specifically, the above-mentioned *Pusarobacter* (Agrobacterium pusense The culture medium for F21 can be LB medium or other conventional media in this field, as long as it can support the growth of *Pusamobacillus* (…). Agrobacterium pusense Any culture medium suitable for the normal growth of F21 can be used. After culturing the strain to the target concentration, treat the fermentation product according to the formulation of the inoculum, following conventional treatment methods in this field.

[0015] Thirdly, the present invention provides the above-mentioned *Pusarobacterium* ( Agrobacterium pusense Application of F21 or microbial agents in any of the following: (1) Degradation of organic pollutants; (2) Prepare products that degrade organic pollutants.

[0016] In some embodiments, the aforementioned organic pollutants include ethyl acetate. Sources of ethyl acetate include, but are not limited to, industrial waste gas, industrial wastewater, contaminated soil, groundwater, and solid waste. Specifically, the application scenarios for the aforementioned strains or microbial agents can include waste gas / wastewater treatment in chemical synthesis and pharmaceutical production processes; waste gas / wastewater treatment in paint, ink, and adhesive production processes; emergency repair of ethyl acetate leaks; biological regeneration and recycling pretreatment of ethyl acetate-containing waste solvents, etc. Regardless of whether it is waste gas, wastewater, waste liquid, soil, or solid waste, the related products prepared using the strains of this invention can degrade ethyl acetate in such scenarios.

[0017] Fourthly, the present invention provides a method for degrading ethyl acetate, comprising: utilizing the above-mentioned *Pseudomonas aeruginosa* (… Agrobacterium pusense F21 or microbial agents can be used to treat environments containing ethyl acetate.

[0018] In some embodiments, the method for degrading ethyl acetate includes: reacting the above-mentioned *Pusarobacter* ( Agrobacterium pusense F21 cells were inoculated into LB liquid medium, cultured, isolated and collected, and then added to the environment to be treated.

[0019] In some embodiments, the initial concentration of ethyl acetate in the environment to be treated is ≤1.44 g / L. It has been verified that the strain of the present invention achieves a degradation rate of 99.17% for 0.36 g / L ethyl acetate in a VOCs sampling bag after 24 h, and a degradation rate of 100.00% for 0.36 g / L ethyl acetate after 48 h. Even at a concentration of 1.26 g / L, ethyl acetate can be significantly removed after 48 h of treatment.

[0020] Whether the bacteria are used to directly treat environments containing ethyl acetate, or the bacteria are prepared into related microbial agents to treat environments containing ethyl acetate, the amount of bacteria or agents added can be adjusted according to the type of environment or the content of ethyl acetate in the environment. This invention does not limit this.

[0021] In some embodiments, the environment to be treated can be one or more of the following: pH 5-10; temperature 20-40°C; NaCl concentration ≤4.0%. Under any one or more of the above conditions, the strain of the present invention can grow normally, thus exhibiting good tolerance to extreme environments and can be applied to ethyl acetate treatment in different scenarios.

[0022] The present invention has the following beneficial effects: The present invention (Pusarnobacterium) Agrobacterium pusense F21, isolated from a deodorization system, is the first strain to be found to efficiently degrade ethyl acetate. It exhibits rapid growth, high viable cell count, and outstanding ethyl acetate removal capabilities. Experiments show that the strain grows most rapidly at 35°C, achieving a high cell density. Furthermore, this strain achieves a 99.17% degradation rate of ethyl acetate at a concentration of 0.36 g / L within 24 hours and 100.00% within 48 hours. Even at ethyl acetate concentrations as high as 1.26 g / L, the strain maintains high removal efficiency, demonstrating excellent substrate removal efficiency. Simultaneously, this strain exhibits good tolerance to extreme environments, growing normally under initial pH conditions of 5.0–10.0, temperatures of 25–40°C, and high salinity conditions of 4.0% NaCl. Therefore, this strain combines highly efficient degradation capabilities with broad environmental adaptability, possessing significant potential for industrial applications. Attached Figure Description

[0023] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0024] Figure 1 This is a colony diagram of *Pusamobacillus* F21 from Example 1; Figure 2 The growth curve of Agrobacterium pusanum-F21 in Example 1; Figure 3 The ethyl acetate removal rate of *Pseudomonas aeruginosa* F21 VOCs test in Example 2; Figure 4The graph shows the growth changes of Agrobacterium pusanum-F21 in Example 3 under different initial pH values; a, b, c, d, e, and f indicate significant differences (p<0.05) in pH, OD600nm, and viable cell count among strains F21; the same letter indicates no significant difference (p>0.05) in viable cell count among strains F21; the letters only represent comparisons between pH (OD600nm or viable cell count); Figure 5 This is a graph showing the growth changes of Agrobacterium pusanum-F21 in Example 4 at different temperatures; a, b, c, and d indicate significant differences (p<0.05) in pH, OD600nm, and viable cell count among strains F21; the same letter indicates no significant difference (p>0.05) in viable cell count among strains F21; the letters only represent pH (OD600nm). 600nm Comparison between (or viable cell count); Figure 6 This is a graph showing the growth changes of Agrobacterium pusanum-F21 under different NaCl concentrations in Example 5; a, b, c, d, and e represent the pH, OD, and OD values ​​of strain F21. 600nm Significant differences were found in viable cell counts among strains F21 (p<0.05); identical letters indicate no significant difference in viable cell counts among strains F21 (p>0.05); letters only represent pH (OD). 600nm Comparison between (or viable cell count); Figure 7 The results of removal rates of different concentrations of ethyl acetate by *Psammonium praecoxibrio* F21 in Example 6 are shown. Detailed Implementation

[0025] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0026] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0027] Example 1 This example uses *Plasnamobacterium* (…). A. pusense Isolation and identification of F21 1. Strains Isolation (1) Take 10.0 g of packing material from a biological trickling filter tower in a biological deodorization treatment system in Hangzhou, add it to a conical flask containing 90 mL of sterile physiological saline and glass beads, and shake at 200 r / min for 1 h at 30℃ to remove the attached substances on the packing material and make the liquid turbid.

[0028] (2) Take 5 mL of supernatant from the conical flask and transfer it into 50 mL of inorganic salt selective medium with ethyl acetate concentration of 25 mg / L. Seal the flask mouth with a silicone stopper, and then add one layer of sealing film and two layers of plastic film to the flask mouth for further sealing.

[0029] (3) The sealed culture medium was placed on a constant temperature shaker at 30℃ and 200 r / min for gradient acclimatization. During the acclimatization process, 5 mL of the culture medium was taken out every day and transferred to 50 mL of fresh culture medium of the same kind. At the same time, the concentration of ethyl acetate was increased to 100.0 mg / L, and acclimatization was stopped.

[0030] (4) After the gradient acclimatization is completed, the final selection medium is diluted with sterile water and spread on LB solid medium. The single colonies that grow are continuously streaked to separate them, and a single strain of bacteria is obtained and named F21.

[0031] The LB medium consisted of: 10.0 g / L tryptone, 5.0 g / L yeast extract, 10.0 g / L sodium chloride, pH 7.0 to pH 7.2, and was sterilized at 121°C for 30 min.

[0032] The inorganic salt selective medium consisted of: 4.72 g potassium nitrate, 0.7 g potassium dihydrogen phosphate, 2.52 g disodium hydrogen phosphate, 0.0244 g magnesium sulfate heptahydrate, and 0.015 g calcium chloride, diluted to 1000 mL with distilled water, pH 7.0, and sterilized at 121℃ for 30 min.

[0033] 2. Species identification of strain F21 (1) Morphological characteristics Colony morphology and microscopic morphology of strain F21 on LB solid medium, as follows: Figure 1 As shown, the colonies are round, yellow on the front, smooth, with neat and clear edges, easy to pick up, and have a short rod-shaped microstructure.

[0034] (2) Gene identification of strain F21 Genomic DNA was extracted from the bacterial strain to be tested using a bacterial DNA extraction kit, following the instructions in the manufacturer's manual. Using the extracted bacterial genomic DNA as a template, 16S rRNA was amplified from strain F21 using the following primers: 27F:5´-AGAGTTTGATCCTGGCTCAG-3´ (SEQ ID NO.1); 1492R:5´-TACGACTTAACCCCAATCGC-3´ (SEQ ID NO.2); After amplification, sequencing yielded the gene sequence shown in SEQ ID NO.3. The assembled sequence was confirmed to belong to [a specific gene group / organization] by BLAST homology sequence search.Agrobacterium pusense This strain was deposited at the China Center for Type Culture Collection on March 4, 2026, at Wuhan University, Wuhan, China, and is classified as *Pusanonobacter*. Agrobacterium pusense F21, accession number: CCTCC NO: M 2026373.

[0035] (3) Growth curve of strain F21 Single colonies of strain F21 were picked and cultured in LB medium for 18 h to obtain fresh seed culture. The inoculum was 2% (20% of the culture volume), and inoculated into LB basal medium. The culture was then incubated in shake flasks at 30℃ and 200 r / min. OD was measured every 2 h. 600nm The growth characteristics of strain F21 were investigated by measuring the number of viable bacteria and the viable count.

[0036] The results are as follows Figure 2 As shown, the bacteria enter the logarithmic growth phase at 8 hours, enter the stationary phase after 18 hours, and reach their maximum viable count of 1.32 × 10⁻⁶ at 20 hours. 10 The concentration of CFU / mL was then reduced to the point of decline, so the subsequent culture time was determined to be 20 h.

[0037] Example 2 This example uses *Plasnamobacterium* (…). A. pusense Growth performance and ethyl acetate removal test of F21 Pseudomonas aeruginosa ( A. pusense F21 cells were cultured in liquid LB seed culture medium at 35°C for 20 h, and the viable cell concentration was 1.32 × 10⁻⁶. 10 CFU / mL. Centrifuge 2.5 mL of bacterial culture at 5000 rpm for 5 minutes, remove the supernatant, resuspend in sterile water, wash, and centrifuge again. Repeat the resuspension and washing process twice. Add the bacterial cells settled at the bottom of the centrifuge tube to a 1L VOCs sampling bag containing 50 mL of inorganic salt selective medium. The formula of this inorganic salt selective medium is: 4.72 g potassium nitrate, 0.7 g potassium dihydrogen phosphate, 2.52 g disodium hydrogen phosphate, 0.0244 g magnesium sulfate heptahydrate, 0.015 g calcium chloride, and distilled water to a final volume of 1000 mL. The pH is 7.0. Sterilize at 121℃ for 30 min.

[0038] Add 20 μl of ethyl acetate to a 1 L VOCs sampling bag; simultaneously set up a blank control with no bacteria and only ethyl acetate and inorganic salt culture medium. Place the 1 L VOCs sampling bag in a constant temperature shaker at 35℃ and 200 r / min for 12 h, 24 h, 36 h, and 48 h. Measure the ethyl acetate content in the control and experimental groups using a portable gas detector. After measuring the ethyl acetate content using the above method, calculate the ethyl acetate removal rate using the following formula.

[0039] Ethyl acetate removal rate =

[0040] The results are as follows Figure 3 As shown, the strain achieved a 97.44% removal rate of ethyl acetate at 12 h. As the strain adapted to ethyl acetate and the concentration of ethyl acetate decreased, the degradation rate began to accelerate. The strain achieved a 99.17% removal rate of ethyl acetate at 24 h and almost completely degraded most of the ethyl acetate at 48 h, reaching a 100.00% removal rate.

[0041] Example 3 This example uses *Plasnamobacterium* (…). A. pusense pH tolerance test of F21 Nine different initial pH values ​​of LB liquid culture medium were prepared, with pH gradients of pH 4.0, pH 5.0, pH 6.0, pH 7.0, pH 8.0, pH 9.0, pH 10.0, pH 11.0, and pH 12.0. A. pusense The growth changes of F21 under different initial pH values ​​are as follows: Figure 4 As shown.

[0042] Figure 4 show A. pusense F21 can grow normally at pH 5.0 to pH 10.0, with a viable count of 2.70 × 10⁻⁶. 9 CFU / mL or higher; the viable count was highest at an initial pH of 7.0, reaching 6.89 × 10⁻⁶. 9 CFU / mL. The viable bacterial count gradually decreases as the pH becomes more acidic or alkaline. When the initial pH is 4.0 or below... A. pusense F21 growth was severely inhibited and unable to occur; when the pH was 11.0 or higher, the growth of the strain was significantly inhibited, falling far below normal levels. These experimental results indicate that... A. pusense F21 has good acid and alkalinity tolerance.

[0043] Example 4 This example uses *Plasnamobacterium* (…). A. pusense F21 temperature tolerance test Seven different incubation temperatures were set: 15℃, 20℃, 25℃, 30℃, 35℃, 40℃, and 45℃. A. pusense F21 seed culture was inoculated into the culture medium at a rate of 1%, and cultured at 35℃ and 200 r / min for 20 h. Samples were taken to measure the pH and OD of each culture medium. 600nm And viable bacterial count, analyze the effects of different temperatures on A. pusense The impact of F21 growth.

[0044] A. pusense The growth changes of F21 at different temperatures are as follows: Figure 5 As shown.

[0045] Figure 5 show A. pusense F21 can grow at temperatures ranging from 20℃ to 40℃, and has a maximum viable count of 1.63 × 10⁻⁶ cells at an ambient temperature of 35℃. 10 CFU / mL; at 20℃, the activity of strain F21 was inhibited, its growth activity was significantly reduced, and the viable count was 6.40 × 10⁻⁶. 8 CFU / mL; at 45℃ and above, bacterial activity is greatly inhibited, and the bacteria cannot grow normally and die.

[0046] Example 5 This example uses *Plasnamobacterium* (…). A. pusense NaCl tolerance test of F21 Five different NaCl concentrations of LB liquid culture medium were prepared: 1.0%, 2.0%, 3.0%, 4.0%, and 5.0%. A. pusense F21 seed culture was inoculated into the culture medium at a rate of 1%, and cultured at 35℃ and 200 r / min for 20 h. Samples were taken to measure the pH and OD of each culture medium. 600nm And viable cell count, analyze the effects of different NaCl concentrations on A. pusense The impact of F21 growth.

[0047] A. pusense The growth changes of F21 under different NaCl concentrations are as follows: Figure 6 As shown.

[0048] Figure 6 show A. pusense F21 is used when the NaCl concentration is 1.0%~4.0%. A. pusense F21 can grow. The viable cell count is highest at a NaCl concentration of 1%, reaching 1.76 × 10⁻⁶. 10 CFU / mL; as the NaCl concentration increases... A. pusenseThe growth rate of F21 begins to decrease. When the NaCl concentration is 1%~3%, the strain grows normally, and the viable count can be maintained at 3.14×10⁻⁶. 9 CFU / mL or higher; when the NaCl concentration was 4%, the growth of the strain was significantly inhibited. When the NaCl concentration was greater than or equal to 5%, the strain essentially ceased to grow. The above experimental results indicate that... A. pusense F21 has a certain degree of osmotic pressure resistance.

[0049] Example 6 This example uses *Plasnamobacterium* (…). A. pusense Test on the removal rate of ethyl acetate of different concentrations by F21 Will A. pusense F21 was cultured in liquid LB seed culture medium at 35°C for 20 h, and its viable cell concentration was 1.32 × 10⁻⁶. 10 CFU / mL. Centrifuge 2.5 mL of bacterial culture at 5000 rpm for 5 minutes, remove the supernatant, resuspend in sterile water, wash, and centrifuge again. Repeat this resuspension and washing process twice. Add the bacterial cells settled at the bottom of the centrifuge tube to a 1L VOCs sampling bag containing 50 mL of inorganic salt selective medium. The formula of this inorganic salt selective medium is: 4.72 g potassium nitrate, 0.7 g potassium dihydrogen phosphate, 2.52 g disodium hydrogen phosphate, 0.0244 g magnesium sulfate heptahydrate, 0.015 g calcium chloride, diluted to 1000 mL with distilled water, pH 7.0, and sterilized at 121℃ for 30 min.

[0050] Ethyl acetate at concentrations of 0.36 g / L, 0.54 g / L, 0.72 g / L, 0.90 g / L, 1.08 g / L, 1.26 g / L, and 1.44 g / L was added to 1L VOCs sampling bags, respectively. A blank control was also established, consisting of culture medium containing only ethyl acetate and inorganic salts without any bacteria. The 1L VOCs sampling bags were incubated in a constant-temperature shaker at 30℃ and 200 r / min for 48 h. The ethyl acetate content in both the control and experimental groups was measured using a portable gas detector.

[0051] The results are as follows Figure 7 As shown, the strain achieved a maximum removal rate of 100.00% after 48 h at an ethyl acetate concentration of 0.36 g / L. At substrate concentrations ranging from 0.36 g / L to 1.08 g / L, the removal rate remained above 95.00% after 48 h. However, at a substrate concentration of 1.44 g / L, the ethyl acetate removal efficiency significantly decreased to 49.73% after 48 h. The concentration of 1.26 g / L represents a relatively high tolerance level in engineering applications, indicating that the strain has good tolerance to ethyl acetate and can treat high-concentration ethyl acetate waste gas and wastewater.

[0052] In summary, the present invention A. pusense F21 can be used for the removal of ethyl acetate, and because it has strong tolerance to high salt and strong acid and alkali conditions, its application scenarios are also more extensive. In addition to the treatment of ethyl acetate in conventional environments, it can also be used for the treatment of ethyl acetate under strong acid and alkali conditions and high concentration conditions, and the effect is stable and the removal rate is high.

[0053] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. Agrobacterium pusa ( Agrobacterium pusense F21, characterized in that, Its accession number is CCTCC NO: M2026373.

2. A microbial inoculant, characterized in that, Contains the *Pusarobacterium* as described in claim 1 ( Agrobacterium pusense )F21.

3. The microbial agent according to claim 2, characterized in that, The microbial agent is selected from any one of liquid formulations, semi-solid formulations, and solid formulations.

4. The microbial agent according to claim 3, characterized in that, The liquid formulation includes the *Pusanobacterium* (…). Agrobacterium pusense F21 fermentation broth, concentrated bacterial suspension, and bacterial cell suspension.

5. The microbial agent according to claim 3, characterized in that, The semi-solid preparations include bacterial mud and bacterial paste.

6. The microbial agent according to claim 3, characterized in that, The solid preparations include bacterial powder and lyophilized powder.

7. The microbial agent according to claim 3, characterized in that, The method for preparing the microbial agent includes: culturing the *Pusarobacterium* as described in claim 1 (…). Agrobacterium pusense )F21.

8. The *Agrobacterium pusa* as described in claim 1 (… Agrobacterium pusense The application of the microbial agent according to any one of claims 2 to 7 (F21) in any of the following: (1) Degradation of ethyl acetate; (2) Prepare products that degrade ethyl acetate.

9. The application according to claim 8, characterized in that, The sources of the ethyl acetate include industrial waste gas, industrial wastewater, contaminated soil, groundwater, and solid waste.

10. A method for degrading ethyl acetate, characterized in that, include: Using the Agrobacterium pusa as described in claim 1 ( Agrobacterium pusense F21 or any one of claims 2 to 7, the microbial agent is used to treat an environment containing ethyl acetate.

11. The method according to claim 10, characterized in that, The initial concentration of ethyl acetate in the environment was ≤1.44 g / L.

12. The method according to claim 11, characterized in that, The environmental conditions are any one or more of the following: pH 5-10; temperature 20-40℃; NaCl concentration ≤4.0%.

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