CN111593009B - Pseudomonas for degrading 3, 4-benzopyrene, method and application - Google Patents
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Abstract
The invention discloses a pseudomonas for degrading 3, 4-benzopyrene, a method and application, and belongs to the technical field of microbiology, biochemistry and fermentation engineering. The Pseudomonas for degrading 3, 4-benzopyrene is Pseudomonas (Pseudomonas sp.) BaP with the preservation number of CCTCC M2020004, and the strain can utilize and degrade 3, 4-benzopyrene and can be used for degradation treatment of 3, 4-benzopyrene pollution in the environment. The pseudomonas is used as a starting bacterium, and the 3, 4-benzopyrene can be degraded by culturing in an inorganic salt liquid culture medium which uses the 3, 4-benzopyrene as a single carbon source. The pseudomonas is cultured in an inorganic salt liquid culture medium which takes 20mg/L of 3, 4-benzopyrene as a single carbon source for 7 days, and the degradation rate of the 3, 4-benzopyrene in the obtained fermentation liquor is about 40 percent.
Description
Technical Field
The invention belongs to the technical field of microbiology, biochemistry and fermentation engineering, and particularly relates to a pseudomonas for degrading 3, 4-benzopyrene, a method and application thereof.
Background
The pseudomonas is aerobic (some pseudomonas can also use nitrate and other nitrogen oxides to perform anaerobic respiration)) Straight or slightly bent gram-negative bacilli, with polar flagella, cannot form spores. Pseudomonas aeruginosa (Pseudomonas aeruginosa) is a major pathogen of nosocomial infections due to its progressive resistance to common antibiotics. Pseudomonas putida (Pseudomonas putida) can be used for degrading toxic garbage (such as aromatic and chlorinated compounds)[1]. Pseudomonas chlororaphis (Pseudomonas chlororaphis) attacks plastics which are useful for waste, especially polyethylene and the like[2]. The 3, 4-benzopyrene is one kind of Polycyclic Aromatic Hydrocarbons (PAHs), and the PAHs pollute the environment, so the PAHs have great harm to the environment because the PAHs have high octanol-water distribution coefficient, belong to fat-soluble substances, have carcinogenic, teratogenic and mutagenic effects on organisms, and can stably exist in the environment. More than half of the 16 PAHs preferentially monitored have carcinogenic effects, and 3, 4-benzopyrene has the highest toxicity. PAHs are not toxic, but can damage biological cell DNA after entering an animal body through a food chain, cause cell DNA chain breakage and change the coding information of genetic materials; the expression transcription level of mitochondrial coding genes can be also adjusted up, and the corresponding antioxidant enzyme activity expression is improved, so that the molecular weight of active oxygen of cells is increased and the cells are oxidized and damaged. With the development of economy, PAHs pollution of different degrees appears in several main economic areas in China, and the pollution of some areas is serious, for example, the total amount of PAHs in the soil of a certain countryside without tin in Jiangsu of Yangtze river Delta is 1058-9500 mu g/kg, the PAHs are mainly 4-6 rings, more than three rings account for 87.0-94.5 percent of the total amount of PAHs, wherein four rings account for 47.9-53.0 percent of the total amount of PAHs[3]. According to the research of the carcinogenic critical concentration of 3, 4-benzopyrene in soil, the critical concentration of the agricultural land is 282 mug/kg[4]According to the existing data, the total amount of PAHs contained in the soil is far higher than the critical value, so that the PAHs in the soil have great carcinogenicity to human bodies.
Various methods for removing polycyclic aromatic hydrocarbons have been proposed, and can be roughly classified into chemical methods, physical methods, and biological methods. The polycyclic aromatic hydrocarbon can be effectively removed by one or a combination of several methods. The following processes are mature: the ultrasonic wave elimination method, the adsorption method, the extraction method, the oxidation method and the ultraviolet light decomposition method have the problems of high cost, complex process, high technical difficulty, serious secondary pollution and the like, and are easy to cause harm to human health and public environment. Accordingly, microbial degradation methods have been proposed. The microbial degradation of PAHs has the characteristics of environmental friendliness, low cost, high efficiency and the like, and becomes an ideal method, an effective means and a main way for eliminating PAHs in the environment, the microbial degradation of PAHs is always a research hotspot at home and abroad, and the screening of functional microorganisms capable of efficiently degrading polycyclic aromatic hydrocarbons is the research core of scientific researchers at present. It is reported that microorganisms capable of degrading polycyclic aromatic hydrocarbons in nature are Bacillus (Bacillus), Pseudomonas (Pseudomonas), Rhodococcus (Rhodococcus), Sphingomonas (Sphingomonas), Flavobacterium (Flavobacterium), and the like. However, the number of high-efficiency degrading strains covered by the method is not large, and particularly, high-ring PAHs degrading bacteria are rare. Therefore, the screening of the high-efficiency degrading bacteria or mixed bacteria of the high-ring PAHs has important practical significance.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a pseudomonas for degrading 3, 4-benzopyrene. The invention also aims to provide application of the pseudomonas and a method for degrading 3, 4-benzopyrene by using the pseudomonas.
The purpose of the invention is realized by the following technical scheme:
in a first aspect, the invention provides a strain of Pseudomonas (BaP strain) for degrading 3, 4-benzopyrene, which is preserved in China center for type culture Collection (preservation date: 2020, 1, 2 and the preservation address: China, Wuhan university), is classified and named as Pseudomonas sp and has the preservation number of CCTCC M2020004. The pseudomonas can utilize and degrade 3, 4-benzopyrene, and can be used for treating 3, 4-benzopyrene pollution in the environment.
In a second aspect, the invention provides an application of the pseudomonas in degrading 3, 4-benzopyrene.
In a third aspect, the invention provides an application of the pseudomonas in degrading benzene, carbazole, benzanthracene, pyrene, benzofluoranthene, phenanthrene and/or dibenzoanthracene.
In a fourth aspect, the invention provides a method for degrading 3, 4-benzopyrene by using the pseudomonas, wherein the pseudomonas is used as a starting bacterium, and the 3, 4-benzopyrene can be utilized and normally grow in an inorganic salt liquid culture medium which takes the 3, 4-benzopyrene as a single carbon source; the method comprises the following steps:
1) the separation of the pseudomonas;
2) identifying the pseudomonas;
3)3, 4-benzopyrene degradation and degradation rate determination:
(3.1) degradation and recovery of 3, 4-benzopyrene:
the culture medium for culturing the pseudomonas is MSM liquid culture medium which takes 3, 4-benzopyrene as a single carbon source;
inoculating the pseudomonas into a test tube containing a liquid LB culture medium, and carrying out shake culture on a shaking table at 30 ℃ for overnight activation;
centrifuging the bacteria liquid after overnight culture, discarding the supernatant, re-suspending with sterile water, centrifuging again, discarding the supernatant, and adding sterile water for re-suspending; the resuspended cell suspension OD600Transferring to a triangular shake flask containing MSM culture medium with 3, 4-benzopyrene as single carbon source, and shake-culturing for 7 d;
thirdly, after culturing for 7 days, adding dichloromethane with the same volume into the triangular shake flask for extraction, pouring the mixture into a separating funnel after shaking extraction of a shaking table, standing, transferring the lower organic phase into a centrifuge tube after liquid is layered, pouring the upper aqueous phase into the triangular flask again, and repeatedly extracting once;
fourthly, centrifuging the extracts obtained by the two-time extraction, and sucking the residual water on the upper layer by a liquid moving device;
fifthly, decompressing and evaporating the extracts obtained in the two times, and adding chromatographic pure acetonitrile into the evaporated round-bottom flask to fully dissolve the 3, 4-benzopyrene;
sixthly, filtering impurities of the obtained acetonitrile solution of the 3, 4-benzopyrene by using an organic filter to prepare a sample to be detected;
(3.2) determination of degradation rate of 3, 4-benzopyrene:
detecting the obtained sample to be detected by using High Performance Liquid Chromatography (HPLC), and calculating the concentration of the sample to be detected according to a standard curve of peak area and concentration which are measured in advance;
detecting by high performance liquid chromatography;
③ the concentration measured by the control group is C1The concentration measured in the experimental group was C2Then, the degradation rate of the final 3, 4-benzopyrene is (C)1-C2)/C1。
Preferably, the MSM liquid culture medium in the step (3.1) contains the following components per liter: KH (Perkin Elmer)2PO4338.8mg,(NH4)2SO4234.0mg,Na2CO3100.0mg，CaCl23.9mg，MgSO4·7H2O 59.3mg，Na2HPO4·12H2O 890.7mg，FeSO40.3mg，FeCl2·4H2O 1.5mg,CoCl2·6H2O 0.19mg,MnSO4·7H2O 0.1mg,ZnCl20.07mg,NiCl2·6H2O 0.024mg，NaMoO4·2H2O0.024mg,MnCl2·4H2O 0.006mg,CuCl2·2H2O 0.002mg。
Further, in the substep (3.2), the type of the used high performance liquid chromatography instrument is Agilent 1100; the column used was an Ultima column of 4.6X 250mm, the mobile phase was acetonitrile, the mobile phase rate was set at 1mL/min, the amount of sample was 30. mu.L per run, and the detection wavelength was 254 nm.
Further, the pH of the liquid medium is 7.0 to 7.2.
Further, the culture conditions are: the culture temperature is 20-30 ℃.
The invention has the following advantages and beneficial effects:
the pseudomonas can be used for degrading 3, 4-benzopyrene through fermentation, the problems of high cost, complex process, high technical difficulty, serious secondary pollution and the like in the traditional PAHs treatment method are avoided, no harm is caused to human health and public environment, and the requirement on experimental conditions is low. The pseudomonas can be preserved permanently, and lays a foundation for mechanism research and further industrial application in the future.
Drawings
FIG. 1 is a phylogenetic tree of this Pseudomonas bacterium (designated BaP3 in the figure);
FIG. 2 is a standard curve of 3, 4-benzopyrene concentration versus peak area;
FIG. 3 shows the degradation efficiency of 3, 4-benzopyrene by Pseudomonas under different 3, 4-benzopyrene concentrations;
FIG. 4 shows the degradation efficiency of 3, 4-benzopyrene by the Pseudomonas under different temperature conditions.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
EXAMPLE 1 isolation, purification and characterization of the Pseudomonas bacterium
The pseudomonas was isolated from the soil of a certain coke plant in the east Steel of Flatstone. The Pseudomonas is preserved in 1 month and 2 days of 2020China center for type culture Collection (Collection Address: Wuhan, China university), named as Pseudomonas by classification sp, preservation number of CCTCC M2020004.The pseudomonas can utilize and degrade 3, 4-benzopyrene.
(1) Isolation of the Pseudomonas bacterium
50g of soil of a certain coking plant of the east Steel of yellow Stone is taken and put into 100mL of inorganic salt liquid culture for shaking culture, and the microorganisms in the soil are enriched. After enrichment culture for one week, taking 5mL of supernatant, transferring the supernatant into 50mL of inorganic salt liquid culture medium which takes 3, 4-benzopyrene as a single carbon source, wherein the concentration of the 3, 4-benzopyrene is 20mg/L, and performing acclimatization culture. Taking culture solution every seven days for subculture, and gradually increasing the concentration of 3, 4-benzopyrene to 40mg/L, 60mg/L and 80 mg/L. And (3) diluting and coating 80mg/L culture solution on an LB solid culture medium, picking single colonies, and carrying out plate streaking separation and purification.
(2) Identification of the Pseudomonas
Taking the pseudomonas, carrying out amplification culture in an LB liquid culture medium, taking fresh bacterial liquid, and extracting the genome of the fresh bacterial liquid by using a bacterial genome extraction kit. Carrying out PCR amplification on the extracted genome by using universal primers 27F and 1492R, wherein the PCR system is as follows: 2 xTaq Plus PCR Master Mix 12.5μL、ddH2O11.5. mu.L, universal primer 27F 0.5. mu.L, universal primer 1492R 0.5. mu.L, and template DNA 0.5. mu.L. And (3) carrying out electrophoresis detection on the product obtained by PCR amplification in 1.5% agarose gel, cutting off a DNA band, and sending the DNA band to a sequencing company for sequencing.
The sequencing results are as follows, with the NCBI GenBank accession number MT539110 for this sequence:
CAGCTACACATGCAGTCGAGCGGATGAGAGGAGCTTGCTCTTCGATTCAGCGGCGGACGGGTGAGTAATGCCTAGGAATCTGCCTAGTAGTGGGGGACAACGTTTCGAAAGGAACGCTAATACCGCATACGTCCTACGGGAGAAAGTGGGGGATCTTCGGACCTCACGCTATTAGATGAGCCTAGGTCGGATTAGCTAGTTGGTAGGGTAAAGGCCTACCAAGGCGACGATCCGTAACTGGTCTGAGAGGATGATCAGTCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGAAAGCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGCCCTCGGGTCGTAAAGCACTTTAAGTTGGGAGGAAGGGCTTACAGCGAATACCTGTGAGTTTTGACGTTACCGACAGAATAAGCACCGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCGCGTAGGTGGCTTGATAAGTTGGATGTGAAATCCCCGGGCTCAACCTGGGAACTGCATCCAAAACTGTCTGGCTAGAGTGCGGTAGAGGGTAGTGGAATTTCCAGTGTAGCGGTGAAATGCGTAGATATTGGAAGGAACACCAGTGGCGAAGGCGACTACCTGGACTGACACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCAACTAGCCGTTGGGATCCTTGAGATCTTAGTGGCGCAGCTAACGCATTAAGTTGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCTGGCCTTGACATGCTGAGAACTTTCCAGAGATGGATTGGTGCCTTCGGGAACTCAGACACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGTAACGAGCGCAACCCTTGTCCTTAGTTACCAGCACGTTATGGTGGGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGGCCAGGGCTACACACGTGCTACAATGGTCGGTACAAAGGGTTGCCAAGCCGCGAGGTGGAGCTAATCCCATAAAACCGATCGTAGTCCGGATCGCAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTAATCGTGAATCAGAACGTCACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGCTCCAGAAGTAGCTAGTCTAACCTTCGGGAGGACGGTACCACGGAG；SEQ ID number NO.1。
Example 23 degradation of 4-benzopyrene and determination of degradation Rate
(1) Degradation and recovery of 3, 4-benzopyrene
Culturing the pseudo sheetThe culture medium of the cytobacteria (Pseudomonas sp.) is MSM liquid culture medium which takes 3, 4-benzopyrene as a single carbon source. MSM liquid culture medium contains the following components per liter: KH (Perkin Elmer)2PO4 338.8mg,(NH4)2SO4 234.0mg,Na2CO3100.0mg，CaCl2 3.9mg，MgSO4·7H2O 59.3mg，Na2HPO4·12H2O 890.7mg，FeSO4 0.3mg，FeCl2·4H2O 1.5mg,CoCl2·6H2O 0.19mg,MnSO4·7H2O 0.1mg,ZnCl2 0.07mg,NiCl2·6H2O 0.024mg，NaMoO4·2H2O 0.024mg,MnCl2·4H2O 0.006mg,CuCl2·2H2O 0.002mg。
1) The Pseudomonas was inoculated into a test tube containing 5mL of liquid LB medium and shake-cultured overnight at 30 ℃ in a shaker.
2) And centrifuging the bacteria liquid after overnight culture at 8000rpm for 2min, discarding the supernatant, resuspending with sterile water, centrifuging at 8000rpm for 2min again, discarding the supernatant, and adding sterile water for resuspension. The resuspended bacterial liquid OD600Adjusted to 1.0, inoculated according to the inoculum size of 2 percent (V/V) to a 250mL triangular shaking flask containing 50mL MSM culture medium taking 3, 4-benzopyrene as a single carbon source, and shake-cultured for 7d at the temperature of 20 ℃ and the speed of 180 rpm.
3) And after culturing for 7 days, adding dichloromethane with the same volume as 50mL into the triangular shake flask for extraction, shaking the table for extraction for 30min, pouring the mixture into a separating funnel, standing for 15min, transferring the lower organic phase into a 50mL centrifuge tube after liquid is layered, pouring the upper aqueous phase into the triangular flask again, and repeatedly extracting once.
4) A total of 100mL of the extract from the two extractions was centrifuged, and the residual water in the upper layer was pipetted off.
5) The extracts obtained in the two times are evaporated to dryness in a water bath at 40 ℃ under reduced pressure by using a rotary evaporator, and 50mL of chromatographically pure acetonitrile is added into a round-bottom flask after the evaporation to dryness so as to fully dissolve the 3, 4-benzopyrene.
6) And (3) filtering out impurities from 1mL of the obtained acetonitrile solution of the 3, 4-benzopyrene by using a 0.45-micrometer organic filter to prepare a sample to be detected.
(2) Determination of degradation rate of 3, 4-benzopyrene
1) The obtained sample to be tested was subjected to HPLC detection, and the concentration was calculated from a calibration curve (FIG. 2) of peak area and concentration measured in advance.
2) And (3) detecting by using a high performance liquid chromatography, wherein the model of the high performance liquid chromatography is Agilent 1100. The column used was an Ultima column of 4.6X 250mm, the mobile phase was acetonitrile, the mobile phase rate was set at 1mL/min, the amount of sample was 30. mu.L per run, and the detection wavelength was 254 nm.
3) The concentration of the control group was C1The concentration measured in the experimental group was C2Then, the degradation rate of the final 3, 4-benzopyrene is (C)1-C2)/C1
Example 3 investigation of the ability of Pseudomonas BaP to degrade other kinds of polycyclic aromatic hydrocarbons
The culture medium for culturing the Pseudomonas sp is MSM liquid culture medium which takes benzene, carbazole, benzanthracene, pyrene, benzofluoranthene, phenanthrene, dibenzanthracene, fluoranthene, naphthalene or ethylbenzene as a single carbon source. MSM liquid culture medium contains the following components per liter: KH (Perkin Elmer)2PO4 338.8mg,(NH4)2SO4 234.0mg,Na2CO3100.0mg，CaCl2 3.9mg，MgSO4·7H2O 59.3mg，Na2HPO4·12H2O 890.7mg，FeSO40.3mg，FeCl2·4H2O 1.5mg,CoCl2·6H2O 0.19mg,MnSO4·7H2O 0.1mg,ZnCl2 0.07mg,NiCl2·6H2O 0.024mg，NaMoO4·2H2O 0.024mg,MnCl2·4H2O 0.006mg,CuCl2·2H2O 0.002mg。
1) The Pseudomonas was inoculated into a test tube containing 5mL of liquid LB medium and shake-cultured overnight at 30 ℃ in a shaker.
2) And centrifuging the bacteria liquid after overnight culture at 8000rpm for 2min, discarding the supernatant, resuspending with sterile water, centrifuging at 8000rpm for 2min again, discarding the supernatant, and adding sterile water for resuspension. The resuspended bacterial liquid OD600Adjusting to 1.0, inoculating to a test tube containing 5mL MSM culture medium with benzene, carbazole, benzanthracene, pyrene, benzofluoranthene, phenanthrene, dibenzoanthracene, fluoranthene, naphthalene or ethylbenzene as single carbon source at an inoculum size of 2% (V/V), and shake culturing at 20 deg.C and 180rpm for 7 d.
3) And observing whether the culture solution obtained after 7d of culture has the growth of the bacteria and the consumption of the polycyclic aromatic hydrocarbon solid substrate.
TABLE 1 degradation of other polycyclic aromatic hydrocarbons by Pseudomonas BaP
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Sequence listing
<110> Wuhan university, Hubei environmental remediation and treatment technology research Limited company
<120> pseudomonas for degrading 3, 4-benzopyrene, method and application
<160> 1
<170> SIPOSequenceListing 1.0
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<211> 1421
<212> DNA
<213> Pseudomonas sp (Pseudomonas sp. BaP)
<400> 1
cagctacaca tgcagtcgag cggatgagag gagcttgctc ttcgattcag cggcggacgg 60
gtgagtaatg cctaggaatc tgcctagtag tgggggacaa cgtttcgaaa ggaacgctaa 120
taccgcatac gtcctacggg agaaagtggg ggatcttcgg acctcacgct attagatgag 180
cctaggtcgg attagctagt tggtagggta aaggcctacc aaggcgacga tccgtaactg 240
gtctgagagg atgatcagtc acactggaac tgagacacgg tccagactcc tacgggaggc 300
agcagtgggg aatattggac aatgggcgaa agcctgatcc agccatgccg cgtgtgtgaa 360
gaaggccctc gggtcgtaaa gcactttaag ttgggaggaa gggcttacag cgaatacctg 420
tgagttttga cgttaccgac agaataagca ccggctaact tcgtgccagc agccgcggta 480
atacgaaggg tgcaagcgtt aatcggaatt actgggcgta aagcgcgcgt aggtggcttg 540
ataagttgga tgtgaaatcc ccgggctcaa cctgggaact gcatccaaaa ctgtctggct 600
agagtgcggt agagggtagt ggaatttcca gtgtagcggt gaaatgcgta gatattggaa 660
ggaacaccag tggcgaaggc gactacctgg actgacactg acactgaggt gcgaaagcgt 720
ggggagcaaa caggattaga taccctggta gtccacgccg taaacgatgt caactagccg 780
ttgggatcct tgagatctta gtggcgcagc taacgcatta agttgaccgc ctggggagta 840
cggccgcaag gttaaaactc aaatgaattg acgggggccc gcacaagcgg tggagcatgt 900
ggtttaattc gaagcaacgc gaagaacctt acctggcctt gacatgctga gaactttcca 960
gagatggatt ggtgccttcg ggaactcaga cacaggtgct gcatggctgt cgtcagctcg 1020
tgtcgtgaga tgttgggtta agtcccgtaa cgagcgcaac ccttgtcctt agttaccagc 1080
acgttatggt gggcactcta aggagactgc cggtgacaaa ccggaggaag gtggggatga 1140
cgtcaagtca tcatggccct tacggccagg gctacacacg tgctacaatg gtcggtacaa 1200
agggttgcca agccgcgagg tggagctaat cccataaaac cgatcgtagt ccggatcgca 1260
gtctgcaact cgactgcgtg aagtcggaat cgctagtaat cgtgaatcag aacgtcacgg 1320
tgaatacgtt cccgggcctt gtacacaccg cccgtcacac catgggagtg ggttgctcca 1380
gaagtagcta gtctaacctt cgggaggacg gtaccacgga g 1421
Claims (4)
1. The pseudomonas capable of efficiently degrading 3, 4-benzopyrene is characterized in that: the Pseudomonas is preserved in China center for type culture Collection, the preservation date is 2020, 01 and 02 months, is classified and named as Pseudomonas sp, and is Pseudomonas BaP with the preservation number of CCTCC M2020004.
2. Use of the pseudomonas strain capable of efficiently degrading 3, 4-benzopyrene according to claim 1 in degrading 3, 4-benzopyrene.
3. Use of the pseudomonas strain capable of efficiently degrading 3, 4-benzopyrene according to claim 1 in degrading benzene, carbazole, benzanthracene, pyrene, benzofluoranthene, phenanthrene and/or dibenzoanthracene.
4. A method for degrading 3, 4-benzopyrene by using the pseudomonas capable of efficiently degrading 3, 4-benzopyrene according to claim 1, wherein the pseudomonas is selected from the group consisting of: culturing the pseudomonas capable of efficiently degrading 3, 4-benzopyrene according to claim 1 in an inorganic salt liquid culture medium using 3, 4-benzopyrene as a single carbon source;
the pH value of the liquid culture medium is 7.0-7.2;
the culture conditions are as follows: the culture temperature is 20-30 ℃.
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| US20040023362A1 (en) * | 2000-02-28 | 2004-02-05 | Grant Stanley | Degradation of polycyclic aromatic hydrocarbons by microorganisms |
| CN104388328A (en) * | 2014-08-26 | 2015-03-04 | 河北农业大学 | New bacterial strain degrading polycyclic aromatic hydrocarbons with five rings or six rings, andacquiring method and application of same |
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| US20040023362A1 (en) * | 2000-02-28 | 2004-02-05 | Grant Stanley | Degradation of polycyclic aromatic hydrocarbons by microorganisms |
| CN104388328A (en) * | 2014-08-26 | 2015-03-04 | 河北农业大学 | New bacterial strain degrading polycyclic aromatic hydrocarbons with five rings or six rings, andacquiring method and application of same |
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| Aromatic compounds lead to increased abundance of antibiotic resistance genes in wastewater treatment bioreactors;Juntao Xia et al.;《Water Research》;20191231;全文 * |
| 苯并芘污染土壤的微生物效应及其降解菌特性的研究;刘君;《中国优秀博硕士学位论文全文数据库（硕士）工程科技Ⅰ辑》;20090215;摘要、第2.2.4.2、3.4.1.4节 * |
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