CN116042444A - PAEs degradation strain fatty bacillus AH-ZY2 and application thereof - Google Patents
PAEs degradation strain fatty bacillus AH-ZY2 and application thereof Download PDFInfo
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- CN116042444A CN116042444A CN202211182628.9A CN202211182628A CN116042444A CN 116042444 A CN116042444 A CN 116042444A CN 202211182628 A CN202211182628 A CN 202211182628A CN 116042444 A CN116042444 A CN 116042444A
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Abstract
The invention relates to a PAEs degradation strain aliphatic bacillus AH-ZY2 and application thereof. The fatty bacillus AH-ZY2 (Pimelobater sp.AH-ZY 2) is preserved in China Center for Type Culture Collection (CCTCC), and is named after classification: pimelobater sp.AH-ZY2, address: mail code 430072 of university of Wuhan in China, with preservation number CCTCC NO: m20221377, storage date 2022, 9 and 5. The beneficial effects of the invention are mainly as follows: the invention provides a novel wild strain capable of degrading various PAEs, which can effectively degrade various PAEs, has high-efficiency degradation capability on short chains and long side chains, and has good application prospects.
Description
Field of the art
The invention relates to a PAEs degradation strain of fatty bacillus AH-ZY2 and application thereof.
(II) background art
Phtalate esters are organic compounds produced by esterification reaction of phthalic anhydride and corresponding alcohols under the action of acid catalyst, are usually colorless transparent oily viscous liquid, are insoluble in water, and are easily dissolved in organic reagents such as methanol, diethyl ether and the like. PAEs can be largely classified into long side chain PAEs (DEHP, dnOP, diNP, etc.) and short side chain PAEs (DEP, DBP, etc.) according to the side chain length. PAEs are mainly added into polyvinyl chloride as a plasticizer to enable the polyvinyl chloride to have elasticity, and are widely used in materials such as plastic products, medical supplies, household appliances and toys. The statistics of the international cancer research Institute (IARC) show that the PAEs content in the finished plastic product is usually 10% -60%, and reports indicate that the content of the mixed PAEs in the plastic can reach up to 70%. The bonding between the PAEs as plasticizers and polymers is not a strong chemical bond, but a physical bond (hydrogen bonding or van der waals forces), so that small changes in environmental factors can accelerate the release of the PAEs from the polymers into the environment. PAEs are a typical environmental endocrine disrupting material, which have hepatorenal toxicity, reproductive toxicity and "three-fold" toxicity to humans. PAEs are stable in environment, difficult to degrade naturally, long in retention time and detected in soil, atmosphere and water, wherein the PAEs in the environment are mainly enriched in the soil. With the mass use of plastic products, the pollution problem of soil by PAEs is more and more paid attention to, wherein DBP, BBP, DEHP, dnOP and the like are listed by the United states Environmental Protection Agency (EPA) as preferential control pollutants, and DEHP and DBP in China are PAEs with highest detection content in the soil.
At present, domestic research on degradation of microbial degradation PAEs mainly focuses on degrading single PAEs, and is usually combined pollution of multiple PAEs in the environment, and strains for efficiently degrading the combined pollution are lacking at present.
(III) summary of the invention
The invention aims to provide a novel wild strain capable of degrading various PAEs, and the strain has a wide substrate spectrum and a good research prospect.
The technical scheme adopted by the invention is as follows:
PAEs degrading bacterium, namely fatty bacillus AH-ZY2 (Pimelobater sp.AH-ZY 2), is preserved in China Center for Type Culture Collection (CCTCC), and is classified and named: pimelobater sp.AH-ZY2, address: mail code 430072 of university of Wuhan in China, with preservation number CCTCC NO: m20221377, storage date 2022, 9 and 5.
The Pimelobater sp.AH-ZY2 was isolated from the farmland soil of the Ubbelopsis.
The 16S rDNA sequence of the strain is shown as SEQ ID No. 1.
SEQ ID No.1：
TTCGGCACTTCGAACGGCTTCCCCCCACAAGGGTTAGGCCACCGGCTTCGGGTGTTGCCGACTTTCGTGACGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCAGCGTTGCTGATCTGCGATTACTAGCGACTCCGACTTCATGGGGTCGAGTTGCAGACCCCAATCCGAACTGAGACCGGCTTTTTGGGATTCGCTCCCCCTCACGGGATCGCAGCCCTTTGTACCGGCCATTGTAGCATGCGTGAAGCCCTGGACATAAGGGGCATGATGACTTGACGTCATCCCCACCTTCCTCCGAGTTGACCCCGGCAGTCTCCCATGAGTCCCCGGCATAACCCGCTGGCAACATGGAACGAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAGCCATGCACCACCTGTACACCAGTATCAAAGAGACCCCTATCTCTAGGGGCTTCCGGTGTATGTCAAACCCAGGTAAGGTTCTTCGCGTTGCATCGAATTAATCCGCATGCTCCGCCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTTAGCCTTGCGGCCGTACTCCCCAGGCGGGGCGCTTAATGCGTTAGCTACGGCACGGAATCCGTGGAAAGGACCCCACACCTAGCGCCCAACGTTTACGGTGTGGACTACCAGGGTATCTAATCCTGTTCGCTCCCCACACTTTCGCTCCTCAGCGTCAGGTAATGCCCAGAGAACCGCCTTCGCCACCGGTGTTCCTCCTGATATCTGCGCATTTCACCGCTACACCAGGAATTCCATTCTCCCCTGCATACCTCTAGTCTGCCCGTATCGAAAGCAAGCAACGAGTTAAGCCCGCTGTTTTCACTCCCGACGCGACAAACCGCCTACGAGCCCTTTACGCCCAATAATTCCGGACAACGCTCGGACCCTACGTATTACCGCGGCTGCTGGCACGTAGTTGGCCGGTCCTTCTTCTGTACCTACCGTCACTTTCGCTTCGTCGGTACTGAAAGAGGTTTACAACCCGAAGGCCGTCATCCCTCACGCGGCGTTGCTGGATCAGGCTTCCGCCCATTGTCCAATATTCCCCACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGACCGGTCACCCTCTCAGGCCGGCTACCCGTCAAAGCCTTGGTGAGCCATTACCTCACCAACAAGCTGATAGGCCGCGAGCACATCCTGGACCGAAAAACTTTCCACCACCACTAGATGCCTAGAGTGGTCATATCCGGTATTAATCACCGTTTCCGGTGGCTATCCCAGAGTCCAGGGCAGATTACTCACGTGTTACTCACCCGTTCGCCGCTCGAGTACCCCCGAAAGGGCCTTCCGCTCGACTGCAGGTAGCAGCGCCAGG
The invention also relates to application of the fatty bacillus AH-ZY2 in microbial degradation treatment of PAEs.
The invention also relates to application of the fatty bacillus AH-ZY2 in repairing soil PAEs pollution.
The PAEs are one or a mixture of more than two of the following: dimethyl phthalate (DMP), diethyl phthalate (DEP), dipropyl phthalate (DPrP), dibutyl phthalate (DBP), butyl Benzyl Phthalate (BBP), diisooctyl phthalate (DEHP), di-n-octyl phthalate (DnOP), diisononyl phthalate (DiNP).
Preferably, the pae is dibutyl phthalate.
The beneficial effects of the invention are mainly as follows: the invention provides a novel wild strain capable of degrading various PAEs, which can effectively degrade various PAEs, has high-efficiency degradation capability on short chains and long side chains, and has good application prospects.
(IV) description of the drawings
FIG. 1 shows the identification result of Pimelobater sp.AH-ZY2 strain; a is colony morphology on LB plate; b is a transmission electron microscope picture; c is the result of gram staining.
FIG. 2 shows the 16S rDNA sequence evolution tree of strain AH-ZY2.
FIG. 3 shows the degradation and growth of various PAEs by strain AH-ZY2.
(fifth) detailed description of the invention
The invention will be further described with reference to the following specific examples, but the scope of the invention is not limited thereto:
example 1:
sample source: the soil isolated from the turnip city, turnip, a. Dove Jiang Oubai, anhui province, was anchored to the farmland soil.
1. Collecting a soil sample: soil samples were taken from the town farmland soil (118°46'35 "E, 31°1' 42" N) anchored by dove Jiang Oubai, turnip, anhui province, and stored in sterile sampling tubes.
2. Screening of PAEs degrading bacteria: taking 5g of soil sample, adding 25ml of water, shaking and mixing uniformly, naturally precipitating for 30min, and respectively sucking 2ml to 50ml of supernatant for culture. After 3 days of incubation, 2mL of the supernatant was added to 100mL of BSM medium (100 mg/L DEP, 100mg/L DBP,100mg/L DEHP,100mg/L DnOP), and incubated at 30℃for 4 days at 180 r/min. Then 2mL of the culture was transferred to a new 100mL BSM medium (100 mg/L DEP, 100mg/L DBP,100mg/L DEHP,100mg/L DnOP were added) and the above-described culture process was repeated. After several times of transfer, the final culture solution is diluted and coated on an LB plate, and single colonies with different morphologies are picked to verify the degradation capacity of the single colonies on PAEs: the purified strain was cultured in BSM medium supplemented with 200mg/L DMP, DEP, DPrP, DBP, BBP, DEHP, dnOP or DiNP, respectively, and specific PAEs were observed for degradation of the strain.
3. Identification of PAEs degrading bacteria: the obtained strain is subjected to physiological and biochemical experimental identification and 16S rDNA sequence sequencing comparison by referring to a Berger system bacteriology manual (2012) to determine the genus of the obtained strain.
4. AH-ZY2 identification results: on LB plate, the colony is white and off-white, the surface is smooth and transparent, the edge is saw-tooth, the whole colony is water stain, and the gram staining result is positive (figure 1).
5. Physiological and biochemical experiments and unique carbon source utilization characteristics: the strain AH-ZY2 was identified by some physiological and biochemical experiments, and the results are shown in Table 1; in addition, the utilization characteristics of the carbon source were examined for 8 unique carbon sources, and the results are shown in Table 2. In tables 1 and 2, "+" indicates that the detection result is positive, and "-" indicates that the detection result is negative.
Table 1: physiological and biochemical experimental results of strain AH-ZY2
Table 2: unique carbon Source utilization Properties of Strain AH-ZY2
Based on the results of the 16S rDNA sequencing alignment (see SEQ ID No.1 for the 16S rDNA sequence of strain AH-ZY 2), an evolutionary analysis was performed, showing that it has the closest affinity to the strain of the genus Pimelobacter (FIG. 2). Therefore, the strain was designated as Pimelobacter sp.AH-ZY2 by combining physiological and biochemical with 16S rDNA identification results.
Example 2:
the PAEs degradation capability test and application experiment of the obtained strain are as follows:
1. the growth was verified with 200mg/L DMP, DEP, DPrP, DBP, BBP, DEHP, dnOP, diNP as the sole carbon source.
2. The strain was tested for degradation over 7 days for 200mg/L DMP, DEP, DPrP, DBP, BBP, DEHP, dnOP, diNP, respectively.
The results of the degradation and growth conditions of the strain AH-ZY2 on the 7 th day are shown in FIG. 3, which shows that the strain AH-ZY2 can effectively degrade all the 8 PAEs tested, can degrade 200mg/L DBP by more than 50% in 7d, and has high-efficiency degradation capability on short chains and long side chains.
Claims (6)
1. PAEs degrading bacterium, namely fatty bacillus AH-ZY2 (Pimelobater sp.AH-ZY 2), is preserved in China Center for Type Culture Collection (CCTCC), and is classified and named: pimelobater sp.AH-ZY2, address: mail code 430072 of university of Wuhan in China, with preservation number CCTCC NO: m20221377, storage date 2022, 9 and 5.
2. The fatty bacillus AH-ZY2 according to claim 1, characterized in that the 16SrDNA sequence of said strain is shown in SEQ ID No. 1.
3. Use of the fatty bacillus AH-ZY2 according to claim 1 for the treatment of PAEs by microbial degradation.
4. Use of the fatty bacillus AH-ZY2 according to claim 1 for repairing soil PAEs pollution.
5. The use according to claim 3 or 4, wherein the PAEs are one or a mixture of two or more of the following: dimethyl phthalate (DMP), diethyl phthalate (DEP), dipropyl phthalate (DPrP), dibutyl phthalate (DBP), butyl Benzyl Phthalate (BBP), diisooctyl phthalate (DEHP), di-n-octyl phthalate (DnOP), diisononyl phthalate (DiNP).
6. The use according to claim 5, wherein the pae is dibutyl phthalate.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211182628.9A CN116042444A (en) | 2022-09-27 | 2022-09-27 | PAEs degradation strain fatty bacillus AH-ZY2 and application thereof |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202211182628.9A CN116042444A (en) | 2022-09-27 | 2022-09-27 | PAEs degradation strain fatty bacillus AH-ZY2 and application thereof |
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- 2022-09-27 CN CN202211182628.9A patent/CN116042444A/en active Pending
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