Bacillus glutamate strain A4 capable of decomposing phthalate and its uses

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 in particular to a method for the decomposition of phthalic acid by a method comprising the steps of: The present invention relates to a Bacillus glutamate A4 strain and uses thereof. [Background technology]

[0002] Phthalic acid esters (PAEs) are the most widely used It is one of the most widely used plasticizers and is one of the most common endocrine disrupting environmental pollutants. One of the most widely studied is phthalates, which are found in plastics, paints, and cosmetics. In 2018, the global production of plastic products was about 300 million This is an increase of 59 million tons, of which 30% comes from China. Only 9% of waste is recycled, and 79% goes directly into landfills or the environment This leads to the rapid accumulation of plastic debris. ) contains a large amount of harmful additives such as plasticizers. Hydrophobicity prevents heavy metals, antibiotics, pesticides and other persistent organic pollutants from entering the micro-plus They adsorb to the surfaces of materials, endangering the soil and causing adverse effects on human health. As the amount of plastic increases, these pollutants affect the ecosystem, such as soil microbial activity and nutrient cycling. PAEs are additives that improve the flexibility of plastic products. PAEs are commonly used plasticizers. However, PAEs are known to be carcinogenic, teratogenic, and mutagenic. If released into the environment, PAEs can inhibit the activity of microorganisms in soil. For example, they can disrupt bacterial communities, reduce urease activity, and enter the food chain through concentration. They are also hazardous to human health. Therefore, PAEs are classified as regulated organic pollutants in many countries. The effects of PAEs on ecosystems and human health have attracted much attention, and many It has become a hot topic in environmental ecology. The decomposition processes of PAEs in the natural environment are mainly hydrolysis, photolysis, and microbial decomposition. In the natural environment, hydrolysis and photolysis are weak, and the decomposition rate is slow. Currently, DMP, DEHP, and DBP Bacterial strains have been isolated that can degrade phthalate pollutants in the environment. However, most of the reported decomposition bacteria are only effective against one type of phthalate ester. It has the ability to efficiently decompose short-chain PAEs (e.g. DMP, DEP, etc.) and long-chain PAEs. There are few reproductive resources of decomposing bacteria that can efficiently decompose both benzophenone and benzoyl peroxidase (e.g., DBP, BBP, etc.). However, it has not been found. Summary of the Invention

[0003] The technical problem that the present invention aims to solve is that most of the reported decomposition bacteria are of one type. It has an efficient decomposition ability only for phthalate esters, and is not effective for short-chain PAEs and long-chain PAEs. Few reproductive resources of decomposing bacteria capable of efficiently degrading both Es have been found. In order to solve the above problems, the technical solutions of the present invention are as follows: The present invention relates to a bacillus subtilis having a phthalic acid decomposition ability. A4 was provided by the China Center for Typical Culture Collection (CCTCC), and its preservation was The storehouse number is CCTCC M20221850. In addition, phthalates include dimethyl terephthalate, diethyl terephthalate, dibutyl terephthalate, butyl terephthalate, and butyl benzyl phthalate. Description: Glutamicibacter sp. A4 is a short-chain glutamic acid bacillus Thalate esters (dimethyl terephthalate, diethyl terephthalate) and long-chain phthalates Effectively decomposes esters (dibutyl terephthalate, butyl benzyl phthalate) can be done. The present invention further provides a use of the Bacillus glutamate A4 strain having the ability to degrade phthalic acid, That is, it is used to prepare a formulation for decomposing phthalate esters. Preferably, the phthalate ester is dimethyl terephthalate and / or diethyl terephthalate. Phthalate and / or Dibutyl Terephthalate and / or Butyl Benzyl Phthalate It is. Description: The formulation for decomposing phthalates is a short-chain phthalate (dimethyl terephthalate). terephthalate, diethyl terephthalate) and long-chain phthalates (dibutyl terephthalate, butyl terephthalate) It can effectively decompose benzoyl phthalate. Preferably, the preparation process of the formulation is as follows: SB1, 500 μL of Bacillus glutamate A4 strain was transferred to 100 ml of LB medium and incubated for 30 After culturing for 24 hours at 150 rpm, the mixture was centrifuged at 8000 rcf for 5 minutes. Obtain a bacterial solution, SB2, bacterial OD after washing the bacterial solution twice with MSM 600 Adjust the value to 1.0 and add the bacterial suspension. The mixture is then temporarily stored at an ambient temperature of 4°C. In the above process, OD 600 The value is the absorbance of a solution at a wavelength of 600 nm. It means. Description: The above uses are for the bacterium Glutamicibacter sp. A Activate 4 for 10-16 h, shake culture until logarithmic phase, collect bacteria, and then use the bacteria as needed. The bacterial content was adjusted to prepare a bacterial suspension, and the prepared bacterial suspension was then soaked in water with a phthalate ester. into contaminated water or soil. Preferably, the process of application of the formulation comprises applying a bacterial suspension to a medium contaminated with phthalates. Put into the body. Description: The application method of the formulation is relatively simple, and the reaction environmental requirements are low. Preferably, the 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% to 20% of the medium mass. Description: Within 72 hours after adding 5% to 20% of the formulation to the medium, four types of phthalate esters were The sterols were almost completely decomposed, and the decomposition rate of all short-chain phthalates exceeded 98%. The degradation rate of long-chain phthalates also reached over 75%, which indicates that the formulation effectively used four types of PAEs. This indicates that the compound has the ability to decompose efficiently. Effect of the Invention

[0004] The present invention has the following beneficial effects: The present invention relates to a bacillus subtilis Glutamicibacter sp. A4 and Bacillus subtilis Glutamic Acid is used in the degradation of phthalate esters. Ibacter sp. A4 was found to be capable of inhibiting dimethyl terephthalate, diethyl terephthalate, Dibutyl terephthalate and butyl benzyl phthalate were used as the sole carbon and energy sources. Under pure culture conditions, the bacterium can grow at 20 mg / kg in mineral salts medium. / L mixed phthalates (Dimethyl terephthalate, Diethyl ... (containing butyl terephthalate, dibutyl terephthalate and butyl benzyl phthalate) It can be almost completely decomposed in 48 hours, with a decomposition rate of over 75%, and the use described in this invention is to prevent environmental pollution. The biological treatment of materials holds wide promise. [Brief description of the drawings]

[0005] [Figure 1] 1 shows the growth morphology of Glutamicibacter sp. A4 of Example 2 when cultured in LB medium for 5 days. [Diagram 2] 1 shows a scanning electron microscope photograph of Glutamicibacter sp. A4 of Example 2. [Diagram 3] FIG. 1 is a schematic diagram showing a phylogenetic tree of 16S rDNA of Glutamicibacter sp. A4 in Example 2. [Figure 4] FIG. 13 is a graph showing the decomposition effect of Glutamicibacter sp. A4 in Example 5 on phthalic acid esters. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0006] In order to make the objectives, technical solutions and advantages of the present invention clearer, reference is made to the accompanying drawings as follows: The present invention will now be described in more detail, but it is apparent that the described embodiments are merely a part of the present invention. The following are merely examples and are not exhaustive. Based on the examples of the present invention, a person skilled in the art may inventively Any other embodiments obtained without any extra labor are within the scope of protection of the present invention. The terms used in the embodiments of the present invention are used only for the purpose of describing specific embodiments, and The term "one kind" used in the examples and claims of the present invention is not intended to limit the scope of the present invention. The singular forms "said" and "the" are used interchangeably unless the context clearly indicates otherwise. It includes numerals and "plurality" generally includes at least two. Example 1 This example describes a phthalic acid decomposition bacillus, Glutamicibacter ... A4 was provided by the China Center for Typical Culture Collection (CCTCC), and The storage number is CCTCC M20221850, and the storage location is Bayi, Wuchang District, Wuhan City, Hubei Province. This is Luohua Coffee Mountain. The glutamic acid bacillus Glutamicibacter sp. A4 was It was isolated from contaminated soil at the Pailou Agricultural University in Kyoto and then concentrated and cultured. The gene sequence of the glutamic acid bacillus Glutamicibacter sp. A4 is shown in SEQ ID NO. GGAGTGGCGGGGTGCTTACACATGCAGTCGAACGATGAAG CCCAGCTTGCTGGGTGGATTAGTGGCGAACGGGTGAGTAA CACGTGAGTAACCTGCCCCCGACTCTGGGATAAGCCCGGG AAACTGGGTCTAATACCGGATATGACCTCGCACCGCATGG TGCGGGGTGGAAAGATTTATCGGTGGGGGATGGACTCGCG GCCTATCAGCTTTGTTGGTGAGGTAATGGCTCACCAAGGCG ACGACGGGTAGCCGGCCTGAGAGGGTGACCGGCCACACTG GGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGT GGGGAATATTGCACAATGGGCGAAAGCCTGATGCAGCGAC GCCGCGTGAGGGATGACGGCCTTCGGGTTGTAACCTCTT TCAGTAGGAAGAAGCGAAAGTGACGGTACCTGCAGAAGA 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 describes a phthalic acid decomposition bacillus, Glutamicibacter ... A4 preparation method is provided for Bacillus glutamate in Example 1. Based on icibacter sp. A4, it includes the following steps: S1, isolating and culturing the strain, including the following steps: S1-1: Approximately 5 g of test soil was added to a 250 mL conical flask, and 100 mL of ultrapure water was added. After that, place the plates in a shaker at 30℃ and 150 rpm, and incubate for 8 hours in the dark. The mixture was allowed to stand for 1 hour, and the resulting supernatant was used as concentrated initial indigenous microorganisms. S1-2: 5 mL of the supernatant was transferred to 95 mL of inorganic salt liquid medium containing phthalic acid ester. The content of phthalate ester in the inorganic salt liquid medium was 5 mg / L, and the culture temperature was 30°C, 150 rpm, and After 5 days of shaking culture in a shaker under the conditions, the inorganic salt liquid medium was The phthalate ester content in the inorganic salt liquid medium was adjusted by continuous enrichment culture, and the above process was carried out. The phthalate ester in the inorganic salt liquid medium was added once, for a total of five times. The content of phthalate esters in the inorganic salt liquid medium was 5 mg / L. After the second addition, The content is 10 mg / L, and the content of phthalate esters in inorganic salt liquid medium after the third addition After the fourth addition, the content of phthalate esters in the inorganic salt liquid medium was 40 mg / L. mg / L, and after the fifth addition, the content of phthalate esters in the inorganic salt liquid medium was 80 mg / L. L, S1-3: Culture solution in inorganic salt liquid medium after five injections for 10 3 Dilute 1:2 and culture the diluted medium. The nutrient solution was applied to an inorganic salt solid medium containing 20 mg / L of phthalate ester, and the medium was incubated at an environmental temperature of 30°C. Cultivate upside down at 37 °C for one day. S1-4: After a single colony grew on solid inorganic salt medium, the single colony was scratched several times. The bacterial strain was isolated and numbered as strain A4, and then the A4 strain was cultured on LB The colony morphology of the A4 strain was observed after transferring the culture medium to a culture medium and culturing it upside down at an environmental temperature of 30°C for 5 days. do. In step S1-2 above, after the first addition, the amount of phthalate ester in the inorganic salt liquid medium The content is 5 mg / L, which means dimethyl terephthalate, diethyl terephthalate, di The content of butyl terephthalate and butyl benzyl phthalate is 5 mg / L. This means that... In step S1-2 above, after the second addition, the phthalate ester in the inorganic salt liquid medium The content of 10 mg / L means dimethyl terephthalate, diethyl terephthalate The content of dibutyl terephthalate and butyl benzyl phthalate is 10 mg / L. This means that. In step S1-2 above, the amount of phthalate ester in the inorganic salt liquid medium after the third addition The content is 20 mg / L, which means dimethyl terephthalate, diethyl terephthalate, The content of dibutyl terephthalate and butyl benzyl phthalate is 20 mg / L. It means something. In step S1-2 above, the amount of phthalate ester in the inorganic salt liquid medium after the fourth addition The content is 40 mg / L, which means dimethyl terephthalate, diethyl terephthalate, The content of dibutyl terephthalate and butyl benzyl phthalate is 40 mg / L. It means something. In step S1-2 above, the amount of phthalate ester in the inorganic salt liquid medium after the fifth addition The content is 80 mg / L, which means dimethyl terephthalate, diethyl terephthalate, The content of dibutyl terephthalate and butyl benzyl phthalate is 80 mg / L. It means something. In this example, the A4 strain was scratch-cultured on LB solid medium for 5 days, and the colonies were pale yellow, Round, mucous, opaque, irregularly shaped edges, raised, moist, smooth surface. , the specific form of which is shown in FIG. S2, identification of strains by scanning electron microscopy, including the following steps: S2-1: A4 strain purified on LB solid medium was inoculated into LB liquid medium and incubated for 10 to 16 h. Transform it into S2-2: 1 mL of bacterial solution was aspirated from the LB liquid medium and centrifuged at 8000 rpm for 3 min. The supernatant was removed, the bacteria were washed three times with sterile MSM medium, and 1 mL of the resulting bacterial pellet was added. Add 2.5% (v / v) glutaraldehyde and mix evenly. Then, leave the mixture at 4°C for 1 hour. Leave it for 0 hours, S2-3: After centrifuging the solution left overnight, the supernatant was poured off and the solution was washed with 0.1M PB, pH 7.0. Wash the bacteria three times with S buffer, each wash lasting 15 min, and finally with 1% osmium. The samples were fixed in acid solution for 1 h. S2-4: Discharge the osmic acid waste and wash the sample with 0.1M, pH 7.0 PBS buffer. The samples were washed three times, each for 15 min, with gradient ethanol solutions. The gradient concentrations were 30%, 50%, 70%, 80%, 90% and 95%. The concentration of each solution was 15 minutes. Dehydrate the samples in 100% ethanol for 20 min. S2-5: The sample was infiltrated with pure acetone for 20 min at a volume ratio of 1 The sample was infiltrated with a mixture of super epoxy resin and acetone in a volume ratio of 3:1 for 1 h, and then diluted with acetone in a volume ratio of 3:1 for 1 h. The sample was infiltrated with a mixture of 1 super epoxy resin and acetone for 3 h, and then soaked in pure su Permeate the samples with epoxy resin for 12 h and encapsulate the permeated samples. , to obtain an encapsulated sample; S2-6: Slice the encapsulated sample with an ultra-thin slicer to a thickness of 70-90 nm. The slices were then soaked in a 1% lead citrate solution and 50% ethylenediaminetetraacetate. The slices were then stained with saturated ethanol for 5 min each, and the stained slices were air-dried and then examined under a transmission electron microscope. Observe the slices in a mirror. In this embodiment, the morphological feature observed by scanning electron microscope is an ellipse. The characteristics are shown in FIG. In the above steps S1 and S2: The inorganic salt liquid medium is an inorganic salt culture medium, and the inorganic salt culture medium component is (NH 4) 2SO4, KH2PO4 at a concentration of 0.5 g / L, and K2HPO4 at a concentration of 1.91 g / L. 3H2O, 0.5g / L NaCl, and 0.2g / L MgSO4 7H2O. The pH value of the inorganic salt culture solution is 7.0. Inorganic salts solid medium is obtained by adding 1.5% (W / V) agar powder to inorganic salts liquid medium. The preparation process of LB liquid medium and LB solid medium is as follows: SA1, 5.0g yeast extract, 10.0g tryptone, 10.0g sodium chloride, ultra-pure Add water to make 1L, adjust the pH to 7.0, and sterilize at 121℃ for 20 minutes to make L. Obtaining B liquid medium, SA2 is based on LB liquid medium and contains 1.5% (W / V) agar powder to create LB solid medium. obtain. S3, identifying the 16S rDNA molecules of the strains, including the following steps: S3-1, total DNA of the strains in the slices was extracted and subjected to bacterial 16S rDNA universal primer The strain genomics was amplified by PCR using primer to obtain PCR products. S3-2: Determine the sequence of the PCR product, and the sequence results were compared with those reported in GenBank. The homology was examined by comparing with the S rDNA sequence, and related species were selected for evolutionary tree analysis and identification. and determine the species of the strain. In the above steps, the results of the evolutionary tree analysis are shown in Figure 3. The 16S rDNA sequence of strain A4 is The sequences were obtained from the NCBI official website (http: / / www.ncbi.nlm.nih The 16S rDNA sequences of other bacterial strains were identified through the BLAST program at The results showed that the strain was most similar to Glutamicibacter sp. The homology was 99%. Therefore, the strains screened in this example were used as Bacillus glutamate (Gl utamicibacter) and Glutamicibacter sp. A It was named 4.

[0008] Example 3 This example describes a phthalic acid decomposition bacillus, Glutamicibacter ... This is a method for preparing C. cerevisiae sp. A4, which differs from Example 1 in the following respects: In step S1-3, the culture solution in the inorganic salt liquid medium after five additions is diluted to 10 4 Diluted twice . In step S1-3, the cells were cultured upside down at an environmental temperature of 30° C. for 2 days. In step S2-1, the A4 strain purified on the LB solid medium was inoculated into the LB liquid medium. Activation was allowed for 13 h. In step S2-2, the mixture was centrifuged at 8000 rpm for 4 minutes and then cooled to 4°C for 1 The mixture was left to stand for 3 h. In step S2-3, the samples were finally fixed in 1% osmic acid solution for 1.5 h. Ta. In step S2-5, the sample was infiltrated with pure super epoxy resin for 14 h. In step S2-6, the slices are soaked in a 1% lead citrate solution and dioxygen acetate. The tissue was stained with 50% ethanol saturated solution for 8 min each.

[0009] Example 4 This example describes a phthalic acid decomposition bacillus, Glutamicibacter ... A method for preparing C. cerevisiae sp. A4 is provided, which differs from Example 1 in the following respects: In step S1-3, the cells were cultured upside down at an environmental temperature of 30° C. for 3 days. In step S2-3, the samples were finally fixed in 1% osmic acid solution for 2 h. In step S2-1, the A4 strain purified on the LB solid medium is inoculated into the LB liquid medium; Activation was allowed for 16 h. In step S2-2, the mixture was centrifuged at 8000 rpm for 5 min and then cooled to 4°C for 1 min. It was left to stand for 6 hours. In step S2-3, the samples were finally fixed in 1% osmic 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 are soaked in a 1% lead citrate solution and dioxygen acetate. The sections were stained with 50% ethanol saturated solution for 10 min each.

[0010] Example 5 This example describes a phthalic acid decomposition bacillus, Glutamicibacter ... The present invention provides a use of Bacillus subtilis sp. A4, and Based on Bacillus sp. A4, A4 was used to prepare a formulation for the degradation of phthalate esters. The preparation process of the formulation is as follows: SB1, 500 μL of Bacillus glutamate A4 strain was transferred to 100 ml of LB medium and incubated for 30 After culturing for 24 hours at 150 rpm, the mixture was centrifuged at 8000 rcf for 5 minutes. Obtain a bacterial solution, SB2, bacterial OD after washing the bacterial solution twice with MSM 600 Adjust the value to 1.0 and add the bacterial suspension. The mixture is then temporarily stored at an ambient temperature of 4°C. This example only shows one method for preparing the preparation, but in practical application, glutamate bacterium can be used. Glutamicibacter sp. A4 was activated for 10-16 h and grown in the logarithmic phase. Culture the bacteria on a shaking rack until 10 min., collect the bacteria, and prepare a bacterial suspension by adjusting the bacterial content according to actual needs. The bacteria suspension was then introduced into water or soil contaminated with phthalates. do. In this example, the method of application of the formulation involves applying the formulation to water contaminated with phthalate esters. The application rate of the formulation is 15% by mass of water. Degradation performance of Glutamicibacter sp. A4: PAEs at a concentration of 20 mg / L (i.e., 20 mg / L DMP, 20 mg / L DEP, 19mL MSM culture containing 20mg / L DBP and 20mg / L BBP Add 1 mL of the bacterial suspension to the liquid, prepare a control group without inoculation, and adjust the pH to 7.0. Each group was replicated three times. The plates were incubated at 30°C and 150 rpm in a thermostatic shaker for 48 hours. Sampling was performed at 3, 6, 12, 24, 48, and 72 hours, and the samples were taken into a conical flask. Add 40 mL of pure methanol for chromatography, shake ultrasonically in a water bath for 1 h, and then sonicate. After completion of the digestion, the mixture was shaken with a vortex mixer, and the supernatant was filtered through a 0.22 μm organic phase filter membrane. The solution was transferred to a 2 mL brown liquid phase vial and detected by high performance liquid chromatography. Chromatography conditions: LC-20AT high-performance liquid phase chromatography (SPD-2A ultraviolet The detection time was 40 min, and the injection volume of the injection system was 20 The separation system used acetonitrile-water as the mobile phase, and the initial flow rate was 1.0 mL / min, PAEs were separated by gradient elution, and the chromatography column was Φ4 .6 x 250mm Inertsil ODS-P liquid chromatography column used The column temperature was 40°C, and the detection system used an ultraviolet detector. m, and detection was performed in dual wavelength detection mode at 225 nm. Degradation effect of Glutamicibacter sp. A4 on four mixed PAEs The results are shown in Figure 4. The bacterium showed a remarkable decomposition effect on four PAEs when cultured with shaking for three days. In particular, the short-chain PAEs (e.g., DMP, DEP) showed a significant increase in the PAEs. In contrast, the decomposition rate of A4 exceeded 95% on the first day and exceeded 98% on the second day. The decomposition rate of AEs (e.g. DBP, BBP) is structurally slow, with DBP decomposition rate reaching 9% on the first day. On the second day, the decomposition rate of BBP reached about 80%. After 72 h of incubation, the four PAEs were almost completely decomposed, and the decomposition rate of short-chain PAEs was The decomposition rate of long-chain PAEs reached more than 75%, which is due to the fact that The ability of Ibacter sp. A4 to efficiently degrade four types of PAEs was confirmed. It shows. In the above performance measurement process, DMP is the abbreviation for phthalate esters, and DEP is the abbreviation for diene. BB is the abbreviation for butyl terephthalate, DBP is the abbreviation for dibutyl terephthalate, and BB is the abbreviation for dibutyl terephthalate. P is the abbreviation for butyl benzyl phthalate.

[0011] Example 6 This example describes a phthalic acid decomposition bacillus, Glutamicibacter ... This is a use of S. cerevisiae sp. A4, which differs from Example 5 in the following respects: The method of application of the formulation is to pour the formulation into soil contaminated with phthalate esters and then apply the formulation. The dosage is 5% of the soil mass. Example 7 This example describes a phthalic acid decomposition bacillus, Glutamicibacter ... This example provides a use of E. coli sp. A4, which differs from Example 5 in the following respects: The method of application of the formulation is to pour the formulation into soil contaminated with phthalate esters and then apply the formulation. The dosage is 20% of the soil mass.

Claims

1. It is preserved at the China Center for Typical Culture Collection (CCTCC), and its preservation number is CCTCC M20. Bacillus glutamate A having a phthalic acid decomposition ability, characterized in that the bacterium is 221850. 4 shares.

2. Phthalates are dimethyl terephthalate, diethyl terephthalate, and dibutyl terephthalate. phthalate, butyl benzyl phthalate, Bacillus glutamate strain A4 having ability to decompose glutamic acid.

3. A use of the Bacillus glutamate A4 strain having the phthalic acid decomposition ability according to claim 1, A use characterized in that it is used for preparing a formulation for decomposing phthalate esters.

4. The phthalate esters are dimethyl terephthalate and / or diethyl terephthalate and and / or dibutyl terephthalate and / or butyl benzyl phthalate, 4. Use according to claim 3.

5. The preparation process of the formulation is as follows: SB1, 500 μL of Bacillus glutamate A4 strain was transferred to 100 ml of LB medium, and incubated for 30 After culturing for 24 hours at 150 rpm at ℃, the mixture was centrifuged at 8000 rcf for 5 minutes. Obtain the bacterial solution, SB2, the bacterial OD after washing the bacterial solution twice with MSM ６００ Adjust the value to 1.0 to prepare and storing the same temporarily at an environmental temperature of 4°C.

6. The method of application of the formulation involves pouring the formulation into a medium contaminated by phthalates.

6. The use according to claim 5, characterized in that

7. 6. The use according to claim 5, characterized in that the medium is water or soil.

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