Bacillus glutamicus A4 strain having phthalic acid-degrading ability and its use

Abstract

The present invention provides a Bacillus glutamate A4 strain capable of decomposing phthalic acid and uses thereof. It belongs to the technical field of biological treatment of environmental pollutants. It is stored at the Center for Typical Culture Collections (CCTCC) under the storage number CCTCC M2022. 1850. Preparation of Bacillus glutamate A4 strain for decomposition of phthalate esters Used in dimethyl terephthalate, diethyl terephthalate, dibutyl terephthalate It has the ability to efficiently decompose butyl benzyl phthalate.

Description

Technical Field

[0001] The present invention relates to the technical field of biological treatment of environmental pollutants, and specifically to Bacillus glutamicum strain A4 having phthalic acid degradation ability and its uses.

Background Art

[0002] Phthalic Acid Esters (PAEs) are one of the most widely used plasticizers and are among the most widely studied environmental pollutants with endocrine disrupting effects. PAEs are widely used in industries such as plastics, paints, and cosmetics. In 2018, the global production volume of plastic products increased by approximately 359 million tons, 30% of which were from China. Of the large amount of plastic waste, only 9% is recycled, and 79% is directly discarded into landfills or the environment, leading to the rapid accumulation of plastic fragments. Microplastics (MPs) contain a large amount of harmful additives such as plasticizers. Due to their large specific surface area and strong hydrophobicity, heavy metals, antibiotics, pesticides, and other persistent organic pollutants adsorb onto the surface of microplastics, endangering the soil and having an adverse impact on human health. With the increase in microplastics, these pollutants affect ecosystem functions such as the activities of soil microorganisms and nutrient cycling. PAEs are additives that improve the flexibility of plastic products and are commonly used plasticizers. However, PAEs are at risk of carcinogenicity, teratogenicity, and mutagenicity. When released into the environment, PAEs inhibit the activities of microorganisms in the soil, for example, interfering with the bacterial community, reducing urease activity, and entering the food chain through concentration. ​​​​​​​​​​​​​​​ Moreover, it threatens human health. Therefore, PAEs are classified as regulated organic pollutants in many countries. The effects of PAEs on the ecosystem and human health have attracted attention and become a hot topic in environmental science and environmental ecology. The decomposition process of PAEs in the natural environment mainly includes hydrolysis, photolysis, and microbial decomposition. However, in the natural environment, hydrolysis and photolysis are weak, and the decomposition rate is slow. Currently, strains capable of decomposing phthalic acid pollutants such as DMP, DEHP, and DBP in the environment have been isolated. However, most of the reported decomposing bacteria have efficient decomposing ability only for one type of phthalic acid ester, and the reproductive resources of decomposing bacteria that can efficiently decompose both short-chain PAEs (such as DMP, DEP, etc.) and long-chain PAEs (such as DBP, BBP, etc.) are hardly found. s (such as DBP, BBP, etc.) are hardly found.

Summary of the Invention

[0003] The technical problem to be solved by the present invention is that most of the reported decomposing bacteria have efficient decomposing ability only for one type of phthalic acid ester, and the reproductive resources of decomposing bacteria that can efficiently decompose both short-chain PAEs and long-chain PAEs are hardly found. Es are hardly found. To solve the above problems, the technical solution of the present invention is as follows: The present invention provides Glutamicibacter sp. A4 with phthalic acid-decomposing ability, which is preserved in the China Center for Type Culture Collection (CCTCC), and its preservation number is CCTCC M20221850. Furthermore, the phthalic acid ester is dimethyl terephthalate, diethyl terephthalate, dibutyl terephthalate, or butyl benzyl phthalate. Description: Glutamicibacter sp. A4 can effectively decompose short-chain phthalic acid esters (dimethyl terephthalate, diethyl terephthalate) and long-chain phthalic acid esters (dibutyl terephthalate, butyl benzyl phthalate). The present invention further provides the use of Glutamicibacter sp. A4 strain having phthalic acid-degrading ability, that is, it is used for the preparation of a formulation for decomposing phthalic acid esters. Preferably, the phthalic acid ester is dimethyl terephthalate and / or diethyl terephthalate and / or dibutyl terephthalate and / or butyl benzyl phthalate. Description: The formulation for decomposing phthalic acid esters can effectively decompose short-chain phthalic acid esters (dimethyl terephthalate, diethyl terephthalate) and long-chain phthalic acid esters (dibutyl terephthalate, butyl benzyl phthalate). Preferably, the preparation process of the formulation is as follows: SB1. Transfer 500 μL of Glutamicibacter sp. A4 strain to 100 ml of LB medium, culture at 30 °C and 150 rpm for 24 h, then centrifuge at 8000 rcf for 5 min to obtain a culture broth, SB2. After washing the broth twice with MSM, adjust the OD value of the bacteria to 1.0 to obtain a bacterial suspension and temporarily store it at an environmental temperature of 4°C. In the above process, the OD value means the absorbance of a certain solution at a wavelength of 600 nm. Description: For the above use, Glutamicibacter sp. A 4 is activated for 10 - 16 h, shaken cultured until the logarithmic phase, the bacteria are collected, and according to actual needs SB3. Centrifuge the bacterial suspension at 8000 rcf for 5 min to obtain a precipitate, and then resuspend the precipitate with MSM to obtain a bacterial suspension with an OD 600 value of 1.0, which is used as a phthalic acid ester-degrading formulation. In the above process, the OD value means the absorbance of a certain solution at a wavelength of 600 nm. 600 In the above use, Glutamicibacter sp. A4 is activated for 10 - 16 h, shaken cultured until the logarithmic phase, the bacteria are collected, and according to actual needs centrifuge the bacterial suspension at 8000 rcf for 5 min to obtain a precipitate, and then resuspend the precipitate with MSM to obtain a bacterial suspension with an OD value of 1.0, which is used as a phthalic acid ester-degrading formulation. activate Glutamicibacter sp. A4 for 10 - 16 h, shake culture until the logarithmic phase, collect the bacteria, and according to actual needs Adjust the bacterial content to prepare a bacterial suspension, and add the prepared bacterial suspension to water or soil contaminated with phthalic acid esters. Preferably, the application process of the formulation is to add the bacterial suspension to a medium contaminated with phthalic acid esters. Description: The application method of the formulation is relatively simple, and the environmental requirements for the reaction are low. Preferably, the above medium is water or soil. Description: The formulation is applicable to both contaminated water and contaminated soil. More preferably, the application amount of the formulation is 5% - 20% of the medium mass. Description: Within 72 hours after adding 5% - 20% of the formulation based on the medium mass, the four phthalic acid esters are almost completely decomposed. The decomposition rate of short-chain phthalic acid esters all exceeds 98%, and the decomposition rate of long-chain phthalic acid esters also reaches 75% or more, indicating that the formulation has the ability to efficiently decompose the four PAEs.

Advantages of the Invention

[0004] The present invention has the following beneficial effects. The present invention provides the use of Glutamicibacter sp. A4 in the decomposition of phthalic acid esters. Glutamicibacter sp. A4 can grow using dimethyl terephthalate, diethyl terephthalate, dibutyl terephthalate, and butyl benzyl phthalate as the sole carbon and energy sources. Under pure culture conditions, this bacterium can decompose 20 mg / L of mixed phthalic acid esters (containing 20 mg / L of dimethyl terephthalate, diethyl terephthalate, dibutyl terephthalate, and butyl benzyl phthalate respectively) in an inorganic salt medium. It can be almost completely decomposed in 48 hours, and the decomposition rate exceeds 75%. The applications described in the present invention are expected to have broad prospects for the biological treatment of environmental pollutants. Substances are expected to have broad prospects.

Brief Description of the Drawings

[0005]

Figure 1

Figure 2

Figure 3

Figure 4

Modes for Carrying Out the Invention

[0006] To make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be described in more detail below with reference to the accompanying drawings. However, it is obvious that the described embodiments are only some Embodiments of the present invention and not all embodiments. Based on the embodiments of the present invention, those skilled in the art can Obtain other embodiments without creative labor, and all are included in the protection scope of the present invention. The terms used in the embodiments of the present invention are only used for the purpose of explaining specific embodiments and do not Limit the present invention. The singular forms such as "one kind", "the above-mentioned" and "said" used in the embodiments and claims of the present invention Include the plural form unless the context clearly indicates otherwise, and "a plurality" generally includes at least two. Example 1 This example is about Glutamicibacter that has the ability to decompose phthalic acid. provided Glutamicibacter sp. A4, which is deposited with the China Center for Type Culture Collection (CCTCC), and its deposit number is CCTCC M20221850, and the deposit location is at Luojia Mountain, Bayi Road, Wuchang District, Wuhan City, Hubei Province. The above-mentioned Glutamicibacter sp. A4 was isolated from the polluted soil of Pailou, Nanjing Agricultural University in Nanjing, and obtained by enrichment culture. The gene sequence of the above-mentioned Glutamicibacter sp. A4 is shown in SEQ ID NO.1, GGAGTGGCGGGGTGCTTACACATGCAGTCGAACGATGAAG CCCAGCTTGCTGGGTGGATTAGTGGCGAACGGGTGAGTAA CACGTGAGTAACCTGCCCCCGACTCTGGGATAAGCCCGGG AAACTGGGTCTAATACCGGATATGACCTCGCACCGCATGG TGCGGGGTGGAAAGATTTATCGGTGGGGGATGGACTCGCG GCCTATCAGCTTGTTGGTGAGGTAATGGCTCACCAAGGCG ACGACGGGTAGCCGGCCTGAGAGGGTGACCGGCCACACTG GGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGT GGGGAATATTGCACAATGGGCGAAAGCCTGATGCAGCGAC GCCGCGTGAGGGATGACGGCCTTCGGGTTGTAAACCTCTT TCAGTAGGGAAGAAGCGAAAGTGACGGTACCTGCAGAAGA AGCGCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGT AGGGCGCAAGCGTTATCCGGATTTATTGGGCGTAAAGAGC​​ TCGTAGGCGGTTTGTCGCGTCTGCCGTGAAAGTCCGAGGC TCAACCTCGGATCTGCGGTGGGTACGGGCAGACTAGAGTG ATGTAGGGGAGACTGGAATTCCTGGTGTAGCGGTGAAATG CGCAGATATCAGGAGGAACACCGATGGCGAAGGCAGGTCT CTGGGCATTTACTGACGCTGAGGAGCGAAAGCATGGGGAG CGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACG TTGGGCACTAGGTGTGGGGGACATTCCACGTTTTCCGCGC CGTAGCTAACGCATTAAGTGCCCCGCCTGGGGAGTACGGC CGCAAGGCTAAAACTCAAAGGAATTGACGGGGGCCCGCAC AAGCGGCGGAGCATGCGGATTAATTCGATGCAACGCGAAG AACCTTACCAAGGCTTGACATGTGCCAGACCGCTTCAGAG ATGGGGTTTCCCTTCGGGGCTGGTTCACAGGTGGTGCATG GTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCC CGCAACGAGCGCAACCCTCGTTCCATGTTGCCAGCACGTA GTGGTGGGGACTCATGGGAGACTGCCGGGGTCAACTCGGA GGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGT CTTGGGCTTCACGCATGCTACAATGGCCGGTACAATGGGT TGCGATACTGTGAGGTGGAGCTAATCCCTAAAAGCCGGTC TCAGTTCGGATTGGGGTCTGCAACTCGACCCCATGAAGTC GGAGTCGCTAGTAATCGCAGATCAGCAACGCTGCGGTGAA TACGTTCCCGGGCCTTGTACACACCGCCCGTCAAGTCACG AAAGTTGGTAACACCCGAAGCCGATGGCCTAACCACCTTG TGTGGGGGGAGTCGTCGAAGTGACGCCGTT。

[0007] Example 2 This example provides a method for preparing Glutamicibacter sp. A4 with phthalic acid degradation ability, based on Glutamicibacter sp. A4 in Example 1, and includes the following steps: Based on Glutamicibacter sp. A4 in Example 1, it includes the following steps: Based on Glutamicibacter sp. A4 in Example 1, it includes the following steps: S1. Isolate and culture the strain, including the following steps: S1-1. Add about 5 g of test soil to a 250 mL conical flask, add 100 mL of ultrapure water, place it on a shaker at 30 °C and 150 rpm, culture it with light shielding for 8 h with shaking, then take it out and let it stand for 2 hours, and use the obtained supernatant as the initial indigenous microorganisms after concentration. hours, and use the obtained supernatant as the initial indigenous microorganisms after concentration. S1-2. Transfer 5 mL of the supernatant to 95 mL of an inorganic salt liquid medium containing phthalic acid ester. The content of phthalic acid ester in the inorganic salt liquid medium is 5 mg / L. After culturing it with shaking on a shaker at 30 °C and 150 rpm for 5 days, with an input amount of 5 mL of the supernatant, the inorganic salt liquid medium is continuously concentrated and cultured to adjust the content of phthalic acid ester in the inorganic salt liquid medium. The above process is taken as one input, and a total of 5 inputs are made. After the first input, the content of phthalic acid ester in the inorganic salt liquid medium is 5 mg / L. After the second input, the content of phthalic acid ester in the inorganic salt liquid medium is 10 mg / L. After the third input, the content of phthalic acid ester in the inorganic salt liquid medium is 10 mg / L. After the third input, the content of phthalic acid ester in the inorganic salt liquid medium is 20 mg / L. After the fourth addition, the content of phthalic acid esters in the inorganic salt liquid medium is 40 mg / L. After the fifth addition, the content of phthalic acid esters in the inorganic salt liquid medium is 80 mg / L, and S1-3. After adding five times, the culture solution in the inorganic salt liquid medium is diluted 10 3 times, and the diluted culture solution is applied to an inorganic salt solid medium with a phthalic acid ester content of 20 mg / L, and cultured by inversion at an environmental temperature of 30°C for 1 day, S1-4. After a single colony grows on the inorganic salt solid medium, the single colony is scraped several times for purification, and the bacterial strain is isolated and numbered as strain A4. Then, strain A4 is transferred to an LB solid medium and cultured by inversion at an environmental temperature of 30°C for 5 days to observe the colony morphology of strain A4. In the above step S1-2, after the first addition, the content of phthalic acid esters in the inorganic salt liquid medium being 5 mg / L means that the contents of dimethyl terephthalate, diethyl terephthalate, di butyl terephthalate, and butyl benzyl phthalate are all 5 mg / L . In the above step S1-2, after the second addition, the content of phthalic acid esters in the inorganic salt liquid medium being 10 mg / L means that the contents of dimethyl terephthalate, diethyl terephthalate , dibutyl terephthalate, and butyl benzyl phthalate are all 10 mg / L . In the above step S1-2, after the third addition, the content of phthalic acid esters in the inorganic salt liquid medium being 20 mg / L means that the contents of dimethyl terephthalate, diethyl terephthalate, dibutyl terephthalate, and butyl benzyl phthalate are all 20 mg / L . ​​In the above step S1-2, the phthalic acid ester in the inorganic salt liquid medium after the fourth addition The content being 40 mg / L means that the contents of dimethyl terephthalate, diethyl terephthalate, dibutyl terephthalate, and butyl benzyl phthalate are all 40 mg / L. This is what it means. In the above step S1-2, the phthalic acid ester in the inorganic salt liquid medium after the fifth addition The content being 80 mg / L means that the contents of dimethyl terephthalate, diethyl terephthalate, dibutyl terephthalate, and butyl benzyl phthalate are all 80 mg / L. This is what it means. In this example, strain A4 was scratch-cultured on an LB solid medium for 5 days, and the colonies were light yellow, circular, mucous, opaque, with an irregular edge, a raised surface, and a wet and smooth surface , and its specific form is shown in Figure 1. S2. Identify the strain by scanning electron microscope observation, including the following steps: S2-1. Inoculate the purified strain A4 on an LB solid medium into an LB liquid medium and activate it for 10-16 h . S2-2. Aspirate 1 mL of the bacterial solution from the LB liquid medium and centrifuge it at 8000 rpm for 3 min , remove the supernatant, add sterile MSM medium to wash the bacteria 3 times, and add 1 mL to the obtained bacterial precipitate of 2.5% (v / v) glutaraldehyde, mix evenly, and let it stand at 4 °C for 1 0 h. S2-3. After centrifuging the solution that has stood overnight, pour off the supernatant, and wash the bacteria 3 times with 0.1 M, pH 7.0 PB S buffer, with each washing time being 15 min. Finally, fix the sample with 1% osmium acid solution for 1 h. S2-4. Pour off the osmium acid waste liquid, and wash the sample with 0.1 M, pH 7.0 PBS buffer Wash three times, with each washing time being 15 min. Treat the sample with ethanol solutions of gradient concentrations to remove water. The gradient concentrations are five kinds of concentrations: 30%, 50%, 70%, 80%, 90% and 95%. The treatment time for each ethanol solution of different concentrations is 15 min. Finally, dehydrate the sample with 100% ethanol for 20 min. S2-5. Immerse the sample in pure acetone for 20 min for infiltration treatment. Immerse the sample in a mixed solution of super epoxy resin and acetone with a volume ratio of 1:1 for 1 h for infiltration, then immerse the sample in a mixed solution of super epoxy resin and acetone with a volume ratio of 3:1 for 3 h for infiltration, and then immerse the sample in pure super epoxy resin for 12 h for infiltration. Encapsulate the infiltrated sample to obtain an encapsulated sample. S2-6. Slice the encapsulated sample with an ultra-thin slicer to obtain slices with a thickness of 70-90 nm. Stain the slices with a 1% lead citrate solution and a 50% ethanol saturated solution of dioxylanil acetate for 5 min each. After air-drying the stained slices, observe the slices with a transmission electron microscope. In this example, the morphological characteristics observed by scanning electron microscope are elliptical, and the specific morphological characteristics are shown in Figure 2. In the above steps S1 and S2: The inorganic salt liquid medium is an inorganic salt culture solution. The components of the inorganic salt culture solution include (NH 4)2SO4 with a concentration of 1.5 g / L, KH2PO4 with a concentration of 0.5 g / L, K2HPO4· 3H2O with a concentration of 1.91 g / L, NaCl with a concentration of 0.5 g / L, and MgSO4·7H2O with a concentration of 0.2 g / L. The pH value of the inorganic salt culture solution is 7.0. The inorganic salt solid medium is obtained by adding 1.5% (W / V) agar powder to the inorganic salt liquid medium. ​​​​​The preparation processes of LB liquid medium and LB solid medium are as follows: SA1. Add ultrapure water to 5.0 g of yeast extract, 10.0 g of tryptone, and 10.0 g of sodium chloride to make up to 1 L, adjust the pH to 7.0, and sterilize at 121 °C for 20 minutes to obtain LB liquid medium. SA2. Based on the LB liquid medium, add 1.5% (W / V) agar powder to obtain LB solid medium. S3. Identify the 16S rDNA molecule of the strain, including the following steps: S3-1. Extract the total DNA of the strain in the slice, and perform PCR amplification of the strain genomics with a bacterial 16S rDNA universal primer to obtain a PCR product. S3-2. Determine the sequence of the PCR product, compare the sequence result with the 16S rDNA sequences reported in GenBank, examine the homology, select the relevant bacterial species for phylogenetic tree analysis and identification, and determine the species of the strain. In the above steps, the result of the phylogenetic tree analysis is shown in Figure 3. The 16S rDNA sequence of strain A4 is compared with the 16S rDNA sequences of other registered bacterial strains through the BLAST program on the NCBI official website (http: / / www.ncbi.nlm.nih.gov / ). As a result, the strain is most similar to Glutamicibacter sp., and the homology rate is 99%. Therefore, the strain screened and obtained in this example was identified as Glutamicibacter, and named Glutamicibacter sp. A4.

[0008] Example 3 This example is about Glutamicibacter with phthalic acid degradation ability. ​​​​​​​​​​​​It is a preparation method of Glutamicibacter sp. A4, which is different from Example 1 in the following aspects: In step S1-3, the culture solution in the inorganic salt liquid medium after 5 additions was diluted 10 4 times. . In step S1-3, it was cultured by inversion for 2 days at an environmental temperature of 30 °C. In step S2-1, the purified A4 strain on the LB solid medium was inoculated into the LB liquid medium and activated for 13 h. In step S2-2, it was centrifuged at 8000 rpm for 4 min and left standing at an environmental temperature of 4 °C for 13 h. In step S2-3, finally, the sample was fixed with a 1% osmium acid solution for 1.5 h . In step S2-5, the sample was infiltrated with pure super epoxy resin for 14 h. In step S2-6, the slices were stained with a 1% lead citrate solution and a 50% ethanol saturated solution of dioxylanil acetate for 8 min each.

[0009] Example 4 This example provides a preparation method of Glutamicibacter sp. A4 with phthalic acid degradation ability, which is different from Example 1 in the following aspects: In step S1-3, it was cultured by inversion for 3 days at an environmental temperature of 30 °C. In step S2-3, finally, the sample was fixed with a 1% osmium acid solution for 2 h. In step S2-1, the purified A4 strain on the LB solid medium was inoculated into the LB liquid medium and activated for 16 h. In step S2-2, it was centrifuged at 8000 rpm for 5 min and left standing at an environmental temperature of 4 °C for 16 h. In step S2-3, finally, the sample was fixed with a 1% osmium acid solution for 2 h. In step S2-3, finally, the sample was fixed with a 1% osmium acid solution for 2 h. In step S2-5, the sample was infiltrated with pure super epoxy resin for 16 h. In step S2-6, the slices were stained with a lead citrate solution with a lead concentration of 1% and a 50% ethanol saturated solution of dioxirane diacetate for 10 min each.

[0010] Example 5 This example provides the use of Glutamicibacter sp. A4 with phthalic acid resolution ability, and based on Glutamicibacter sp. A4 in Example 1, Glutamicibacter sp. A4 was used in the preparation of a formulation for phthalate ester decomposition. The preparation process of the formulation is as follows: SB1. Transfer 500 μL of Glutamicibacter A4 strain to 100 ml of LB medium, and culture it at 30 °C and 150 rpm for 24 h, then centrifuge at 8000 rcf for 5 min to obtain the culture bacterial liquid. SB2. After washing the bacterial liquid twice with MSM, adjust the OD value of the bacteria to 1.0 to obtain a bacterial suspension and temporarily store it at an environmental temperature of 4 °C. This example only shows one preparation method of the formulation. In actual applications, Glutamicibacter 600 sp. A4 is activated for 10 - 16 h, shaken and cultured until the logarithmic phase, the bacteria are collected, and the bacterial content is adjusted according to actual needs to prepare a bacterial suspension, and the prepared bacterial suspension is added to water or soil contaminated with phthalate ester. In this example, the application method of the formulation is to apply the formulation to water contaminated with phthalate ester, and the application amount of the formulation is 15% of the mass of the water. Measurement of the decomposition performance of Glutamicibacter sp. A4 bacteria: ​ 19 mL of MSM culture solution containing PAEs at a concentration of 20 mg / L (i.e., containing 20 mg / L DMP, 20 mg / L DEP, 20 mg / L DBP, and 20 mg / L BBP) was inoculated with 1 mL of the above bacterial suspension, and a control group without inoculation was prepared. The pH was adjusted to 7.0, and each group was repeated three times. The cultures were incubated at 30 °C and 150 rpm in a constant temperature shaker for 48 h, and samples were taken at 1, 3, 6, 12, 24, 48, and 72 h, respectively. 40 mL of pure methanol for chromatography was added to the taken conical flask, and ultrasonic oscillation was performed in a water bath for 1 h. After the ultrasonic treatment was completed, the mixture was vortexed and then the supernatant was filtered through a 0.22 μm organic phase filter membrane, transferred to a 2 mL brown liquid phase vial, and detected by high performance liquid chromatography. Chromatography conditions: LC-20AT high performance liquid chromatography (equipped with an SPD-2A ultraviolet line detector) was used. The detection time was 40 min, the injection volume of the injection system was 20 μL, the separation system used acetonitrile-water as the mobile phase, the initial flow rate was 1.0 mL / min, and PAEs were separated by gradient elution. The chromatography column used was a Φ4 .6 × 250 mm Inertsil ODS-P liquid chromatography column, the column temperature was 40 °C, the detection system used an ultraviolet detector, and detection was performed in a two-wavelength detection mode at 205 nm and 225 nm, respectively. The degradation effect of Glutamicibacter sp. A4 bacteria on four mixed PAEs is shown in Figure 4. This bacterium showed a significant degradation effect on four PAEs under shaking culture for 3 days. In particular, for short-chain PAEs (such as DMP and DEP), the degradation rate of A4 on the above PAEs on the first day exceeded 95%, and the degradation rate on the second day exceeded 98%. For long-chain P ​​​​The decomposition rate of AEs (e.g., DBP, BBP) is inherently slow. The decomposition rate of DBP reached 9% on the first day, and that of BBP was about 70%. However, on the second day, the decomposition rate of BBP reached about 80%. After 72 hours of cultivation, the four PAEs were almost completely decomposed. The decomposition rate of short-chain PAEs exceeded 98%, and that of long-chain PAEs also reached over 75%, indicating that Glutamicibacter sp. A4 has the ability to efficiently decompose the four PAEs. In the above performance measurement process, DMP is the abbreviation of phthalic acid ester, DEP is the abbreviation of diethyl terephthalate, DBP is the abbreviation of dibutyl terephthalate, and BBP is the abbreviation of butyl benzyl phthalate. Example 6 This example is about the use of Glutamicibacter sp. A4 with phthalic acid degradation ability, which is different from Example 5 in the following aspects: The application method of the formulation is to put the formulation into the soil contaminated by phthalic acid esters, and the application amount of the formulation is 5% of the soil mass. Example 7 This example provides the use of Glutamicibacter sp. A4 with phthalic acid degradation ability, which is different from Example 5 in the following aspects: The application method of the formulation is to put the formulation into the soil contaminated by phthalic acid esters, and the application amount of the formulation is 20% of the soil mass.

[0011] ​​​​​​​​​​

Claims

1. It is deposited in the China Center for Type Culture Collection (CCTCC), and its deposit number is CCTCC M20 221850, and it is characterized by having the ability to decompose phthalic acid, Bacillus glutamate A 4 strains.

2. The phthalic acid ester is dimethyl terephthalate, diethyl terephthalate, dibutyl tere phthalate or butyl benzyl phthalate, and it is characterized by the phthalic acid decomposition ability described in Claim 1 Bacillus glutamate A4 strains.

Images