CN112940958A - Strain capable of effectively degrading short-chain chlorinated paraffin, and rapid screening method and application thereof - Google Patents
Strain capable of effectively degrading short-chain chlorinated paraffin, and rapid screening method and application thereof Download PDFInfo
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- 238000012216 screening Methods 0.000 title claims abstract description 14
- 241000588724 Escherichia coli Species 0.000 claims abstract description 23
- 230000015556 catabolic process Effects 0.000 claims abstract description 22
- 238000006731 degradation reaction Methods 0.000 claims abstract description 22
- 241000894006 Bacteria Species 0.000 claims abstract description 9
- 239000010802 sludge Substances 0.000 claims abstract description 7
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- 239000001301 oxygen Substances 0.000 claims abstract description 6
- 229910052760 oxygen Inorganic materials 0.000 claims abstract description 6
- 230000001580 bacterial effect Effects 0.000 claims abstract description 5
- 239000002609 medium Substances 0.000 claims description 24
- 239000001963 growth medium Substances 0.000 claims description 21
- 239000001888 Peptone Substances 0.000 claims description 15
- 108010080698 Peptones Proteins 0.000 claims description 15
- 235000015278 beef Nutrition 0.000 claims description 15
- 235000019319 peptone Nutrition 0.000 claims description 15
- 239000007787 solid Substances 0.000 claims description 15
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 7
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 6
- 229910052799 carbon Inorganic materials 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 6
- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
- 229910052698 phosphorus Inorganic materials 0.000 claims description 6
- 239000011574 phosphorus Substances 0.000 claims description 6
- 239000011573 trace mineral Substances 0.000 claims description 6
- 235000013619 trace mineral Nutrition 0.000 claims description 6
- 239000000706 filtrate Substances 0.000 claims description 4
- 108090000623 proteins and genes Proteins 0.000 claims description 4
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 3
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 3
- 229910000366 copper(II) sulfate Inorganic materials 0.000 claims description 3
- 238000002474 experimental method Methods 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
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- 238000002360 preparation method Methods 0.000 claims description 3
- 238000006748 scratching Methods 0.000 claims description 3
- 230000002393 scratching effect Effects 0.000 claims description 3
- 239000001632 sodium acetate Substances 0.000 claims description 3
- 235000017281 sodium acetate Nutrition 0.000 claims description 3
- 230000000694 effects Effects 0.000 abstract description 9
- 239000010865 sewage Substances 0.000 abstract description 4
- 230000008878 coupling Effects 0.000 abstract description 2
- 238000010168 coupling process Methods 0.000 abstract description 2
- 238000005859 coupling reaction Methods 0.000 abstract description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 6
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- 230000007613 environmental effect Effects 0.000 description 3
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- 238000004321 preservation Methods 0.000 description 3
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- 238000011160 research Methods 0.000 description 3
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- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 241000589516 Pseudomonas Species 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 2
- 239000012620 biological material Substances 0.000 description 2
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- 230000036541 health Effects 0.000 description 2
- 239000004021 humic acid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- RNFJDJUURJAICM-UHFFFAOYSA-N 2,2,4,4,6,6-hexaphenoxy-1,3,5-triaza-2$l^{5},4$l^{5},6$l^{5}-triphosphacyclohexa-1,3,5-triene Chemical compound N=1P(OC=2C=CC=CC=2)(OC=2C=CC=CC=2)=NP(OC=2C=CC=CC=2)(OC=2C=CC=CC=2)=NP=1(OC=1C=CC=CC=1)OC1=CC=CC=C1 RNFJDJUURJAICM-UHFFFAOYSA-N 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 229910003321 CoFe Inorganic materials 0.000 description 1
- 241000588722 Escherichia Species 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 206010064571 Gene mutation Diseases 0.000 description 1
- 241000316848 Rhodococcus <scale insect> Species 0.000 description 1
- 208000031320 Teratogenesis Diseases 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
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- 150000001335 aliphatic alkanes Chemical class 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000005571 anion exchange chromatography Methods 0.000 description 1
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- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 229910052753 mercury Inorganic materials 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
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- 231100000956 nontoxicity Toxicity 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
- 238000013033 photocatalytic degradation reaction Methods 0.000 description 1
- 239000004014 plasticizer Substances 0.000 description 1
- 239000010801 sewage sludge Substances 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
- 239000004408 titanium dioxide Substances 0.000 description 1
- 238000004065 wastewater treatment Methods 0.000 description 1
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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Abstract
A bacterial strain for effectively degrading short-chain chlorinated paraffin, a rapid screening method and application thereof are characterized in that: the strain is SCCPs degrading bacteria, and is named in classification: escherichia coli with a deposit number of: CGMCC No. 20989. The method for coupling pressurized dissolved oxygen with large-dose addition of SCCPs is used for the first time, and an Escherichia coli strain is rapidly screened from aerobic activated sludge of a sewage treatment plant, has a good degradation and removal effect on the SCCPs, and can degrade the SCCPs to a lower degree within 14 days; the degradation and removal effect of the Escherichia coli strain on the SCCPs is realized for the first time.
Description
Technical Field
The invention relates to the technical field of industrial wastewater treatment, in particular to a strain for effectively degrading short-chain chlorinated paraffins (SCCPs) and a rapid (less than 5 days) screening method and application thereof. The strain is already preserved in 2020.11.02 in China general microbiological culture Collection center, and the preservation numbers are: cgmccno.20989, biological material (strain) deposited: SCCPs degrading bacteria, classified name: escherichia coli (Escherichia coli).
Background
Chlorinated paraffin is widely used as an additive of metal cutting fluid in metal processing, a plasticizer in plastic processing, and a flame retardant for paint, textiles and the like. The long-chain, medium-chain and short-chain chlorinated paraffin can be divided according to the length of a molecular chain, the short-chain chlorinated paraffin (SCCPs) has the most special physicochemical properties compared with the medium-chain and long-chain chlorinated paraffins, has the 'three-cause effect' of carcinogenesis, teratogenesis and gene mutation, and is finally listed in 'St Golgi convention about persistent organic pollutants' annex A controlled Persistent Organic Pollutants (POPs) list in 2017.
SCCPs have not heretofore been found to be of natural origin, primarily from human activity, through point-source, non-point-source discharge (including accidental spillage or flushing, etc.) into environmental recipients during production, storage, transportation, product manufacture, and use of chlorinated paraffins. Meanwhile, although most of SCCPs can be adsorbed and removed by sludge in the traditional sewage treatment plant, part of SCCPs can enter a receiving water body, or be adsorbed on suspended particles, or sink into water body bottom mud, or volatilize into the atmosphere, and are adsorbed by atmospheric particulate matters, so that the atmospheric long-distance transportation is realized. In addition, SCCPs have strong biological enrichment and biological amplification, are detected in many marine products in offshore sea areas in China, and seriously threaten environmental safety and human health. Therefore, the research on the efficient SCCPs removal method has higher economic and social values for improving the quality of marine products and ensuring environmental safety and human health.
At present, however, research on removal methods of SCCPs is not common at home and abroad, and removal methods appearing in literature mainly include zero-valent iron reduction, photocatalytic degradation, and microbial degradation removal. Such as Zhang, etc. by using nano zero-valent iron particles to reduce and dechlorinate to remove SCCPs. Research shows that the dechlorination rate of the SCCPs by the nano zero-valent iron is related to the pH value, the adding amount, the temperature and the adding amount of humic acid. Proper humic acid is added to contribute to the reduction of the SCCPs by the nano zero-valent iron, but when the concentration is higher than 15mg/L, the dechlorination rate is inhibited. Koh et al found that SCCPs can be effectively degraded by the synergistic effect of an ultraviolet lamp (medium-pressure mercury lamp) and hydrogen peroxide, and the degradation product has no toxicity to microorganisms basically after photocatalytic treatment for 300 min. On the basis, Chen et al synthesized a redox graphene (RGO/CoFe)2O4Ag) photocatalytic material, can be used for degrading SCCPs under visible light and is matched with commercial titanium dioxide P25TiO2A comparison was made. The result shows that the catalyst can remove 91.9 percent of SCCPs in 12h, which is far higher than that of P25TiO221.7% of. However, the high cost of the physical and chemical method limits the technical popularization and application, and the microbiological method is favored by researchers as the most economic method. Lu screens out a strain of pseudomonas N35 (gram-negative bacteria), which can take SCCPs as a carbon source and an energy source, can effectively degrade the SCCPs and can dechlorinate 57.5% in 20 days. The bacteria added into the sewage sludge can remove 73.4 percent of SCCPs, but the degradation period is longer and needs 30 days. Heath et al also screened a strain of Pseudomonas 273 which had a good dechlorination effect on chlorinated alkanes, but like the Lu-screened strain, the bacterial dechlorination cycle was longer, requiring 20 days or even longer. In addition to gram-negative bacteria, gram-positive bacteria also have a degrading effect on SCCPs. For example, Allpress et al screened a gram-positive strain Rhodococcus sp S45-1, which was also identifiedSCCPs are the only carbon source and energy source for metabolic degradation, and the only defect is that the degradation time is long and needs 30-100 days. Although previous studies have shown that either gram-positive or gram-negative bacteria have a degrading effect on SCCPs, the biodegradation cycles are often long. Therefore, it is of great significance to find strains capable of efficiently degrading SCCPs.
Disclosure of Invention
Aiming at the defects in the prior art, the strain capable of effectively degrading the SCCPs to a lower degree is screened out within 14 days, and the strain rapid screening method is provided.
In order to solve the technical problem, the technical scheme adopted by the application is as follows: a bacterial strain for effectively degrading short-chain chlorinated paraffin is SCCPs degrading bacteria, and is classified and named as: escherichia coli (Escherichia coli) with a deposit number of: CGMCC No.20989, the preservation address is the institute of microbiology of China academy of sciences No. 3, Xilu No.1, Beijing, Chaoyang.
The application also provides a sequence number of the strain for effectively degrading the short-chain chlorinated paraffin, and specifically refers to a gene sequence shown in SEQ ID NO. 1.
The application also provides application of the strain in degradation of short-chain chlorinated paraffin.
The application also provides a screening method of the strain for effectively degrading the short-chain chlorinated paraffin, which comprises the following specific steps:
(1) adopting 1000mg/L sodium acetate as a carbon source of a culture medium, and simultaneously adding a nitrogen source, a phosphorus source and trace elements;
(2) filtering activated sludge, adding the filtrate into a pressurized dissolved oxygen reactor, adding SCCPs with the final concentration of 50-200mg/L, adjusting the pressure to 0.3MPa by oxygenation, and performing acclimation in the culture medium in the step (1) for 2-3 d; then carrying out plate streaking on a beef extract peptone solid culture medium, and selecting strains which can grow out on the culture medium;
(3) re-screening of strains: and (2) selecting a strain which can grow out of the beef extract peptone solid medium into a liquid medium, wherein the liquid medium is the same as an acclimatization medium (the acclimatization medium is a culture medium which contains a carbon source, a nitrogen source, a phosphorus source and trace elements in the step (1)), adding SCCPs with the concentration of 20mg/L, and performing an SCCPs degradation experiment.
Preferably, the concentrations of the nitrogen source, the phosphorus source and the trace elements added in the step (1) are as follows: 825mg/L (NH)4)2SO4，340mg/L KH2PO4，10mg/L MgSO4，20mg/L Fe2(SO4)3，4mg/L ZnSO4，3.2mg/L MnSO4，1mg/L CuSO4(i.e., the final concentration of each component in the medium after addition).
Preferably, the specific preparation process of the beef extract peptone solid medium in the step (2) comprises the following steps: adding SCCPs with the final concentration of 50mg/L into a beef extract peptone solid culture medium, then carrying out autoclaving at 121 ℃ for 30min, subpackaging the culture medium and pouring into a culture dish, and scratching a plate after the culture medium is solidified; after autoclaving, beef extract peptone medium is filtered and added with SCCPs. Because this would cause air bubbles in the solid medium during shaking, which would be detrimental to plate streaking of subsequent strains.
The application has the advantages and beneficial effects that:
1. the method for coupling pressurized dissolved oxygen with large-dose addition of SCCPs is used for the first time, and an Escherichia coli strain is rapidly screened from aerobic activated sludge of a sewage treatment plant, has a good degradation and removal effect on the SCCPs, and can degrade the SCCPs to a lower degree within 14 days; the degradation and removal effect of the Escherichia coli strain on the SCCPs is realized for the first time.
2. After the strain is used for 14 days for degrading SCCPs, the removal rate of the SCCPs can reach 89.88%, and Cl in solution is removed from the SCCPs-The content is 11.71 mg/L; greatly shortens the degradation period and time and improves the treatment efficiency.
Drawings
FIG. 1 shows Escherichia coli (Escherichia coli strain) selected.
FIG. 2 changes in SCCPs before and after 14 days of Escherichia coli degradation.
FIG. 3 changes in chloride ion before and after 14 days of Escherichia coli degradation of SCCPs.
Detailed Description
The present application is described in further detail below by way of specific embodiments, but the present application is not limited to only the following examples.
Examples
The specific strains of the present application were screened and prepared as follows:
(1) firstly, adding a filtrate of aerobic activated sludge of a sewage treatment plant into a pressurized dissolved oxygen reaction device, and screening and domesticating by using SCCPs with certain concentration, wherein the specific method comprises the following steps: the carbon source of the culture medium is 1000mg/L sodium acetate, and simultaneously, a proper amount of nitrogen source, phosphorus source and trace elements are added, wherein the final concentration of each element in the culture medium is as follows: 825mg/L (NH)4)2SO4，340mg/L KH2PO4，10mg/L MgSO4，20mg/L Fe2(SO4)3，4mg/L ZnSO4，3.2mg/L MnSO4，1mg/L CuSO4；
(2) Filtering activated sludge through filter paper, adding the obtained filtrate into a pressurized dissolved oxygen reactor, adding a large amount of SCCPs with the concentration of 50-200mg/L, adjusting the pressure to 0.3MPa through oxygenation, and performing acclimation for 2-3d in the culture medium prepared in the step (1); then, carrying out plate streaking on a beef extract peptone solid medium (the medium contains 50mg/LSCCPs), and selecting strains (2-3 colonies) which can grow on the beef extract peptone solid medium;
preparation of beef extract peptone solid Medium containing 50mg/L SCCPs: adding SCCPs with the final concentration of 50mg/L into a beef extract peptone solid culture medium, then carrying out autoclaving at 121 ℃ for 30min, subpackaging the culture medium and pouring into a culture dish, and scratching a plate after the culture medium is solidified; after autoclaving, beef extract peptone medium is filtered and added with SCCPs. Because this would cause air bubbles in the solid medium during shaking, which would be detrimental to plate streaking of subsequent strains.
(2) Re-screening of strains: selecting strains capable of growing in a beef extract peptone solid medium, introducing the strains into a liquid medium, wherein the liquid medium is the same as the domestication medium, adding SCCPs with the concentration of 20mg/L, and performing an SCCPs degradation experiment.
(3) The selected strain is shown in the attached figure 1, and is selected Escherichia coli strain with the preservation number: CGMCC No.20989, biological material (strain) preserved: SCCPs degrading bacteria, classified name: escherichia coli (Escherichia coli);
the specific gene sequence is as follows:
TCAGATTGAACGCTGGCGGCAGGCCTAACACATGCAAGTCGAACGGTAACAGGAA GAAGCTTGCTTCTTTGCTGACGAGTGGCGGACGGGTGAGTAATGTCTGGGAAACT GCCTGATGGAGGGGGATAACTACTGGAAACGGTAGCTAATACCGCATAACGTCGCA AGACCAAAGAGGGGGACCTTCGGGCCTCTTGCCATCGGATGTGCCCAGATGGGAT TAGCTAGTAGGTGGGGTAACGGCTCACCTAGGCGACGATCCCTAGCTGGTCTGAG AGGATGACCAGCCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCA GCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCCATGCCGCGTGTAT GAAGAAGGCCTTCGGGTTGTAAAGTACTTTCAGCGGGGAGGAAGGGAGTAAAGTT AATACCTTTGCTCATTGACGTTACCCGCAGAAGAAGCACCGGCTAACTCCGTGCCA GCAGCCGCGGTAATACGGAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAG CGCACGCAGGCGGTTTGTTAAGTCAGATGTGAAATCCCCGGGCTCAACCTGGGAA CTGCATCTGATACTGGCAAGCTTGAGTCTCGTAGAGGGGGGTAGAATTCCAGGTGT AGCGGTGAAATGCGTAGAGATCTGGAGGAATACCGGTGGCGAAGGCGGCCCCCTG GACGAAGACTGACGCTCAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATAC CCTGGTAGTCCACGCCGTAAACGATGTCGACTTGGAGGTTGTGCCCTTGAGGCGT GGCTTCCGGAGCTAACGCGTTAAGTCGACCGCCTGGGGAGTACGGCCGCAAGGTT AAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAAT TCGATGCAACGCGAAGAACCTTACCTGGTCTTGACATCCACAGAACTTTCCAGAG ATGGATTGGTGCCTTCGGGAACTGTGAGACAGGTGCTGCATGGCTGTCGTCAGCTC GTGTTGTGAAATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATCCTTTGTTG CCAGCGGTCCGGCCGGGAACTCAAAGGAGACTGCCAGTGATAAACTGGAGGAAG GTGGGGATGACGTCAAGTCATCATGGCCCTTACGACCAGGGCTACACACGTGCTAC AATGGCGCATACAAAGAGAAGCGACCTCGCGAGAGCAAGCGGACCTCATAAAGT GCGTCGTAGTCCGGATTGGAGTCTGCAACTCGACTCCATGAAGTCGGAATCGCTAG TAATCGTGGATCAGAATGCCACGGTGAATACGTTCCCGGGCCTTGTACACACCGCC CGTCACACCATGGGAGTGGGTTGCAAAAGAAGTAGGTAGCTTAACCTTCGGGAGG GCGCTTACCACTTTGTGATTCATGACTGGGGTGAAGT。
the specific degradation experimental conditions and procedures are as follows:
gas chromatography conditions (GC 2030-ECD):
the type of the chromatographic column: inertcap5, 30m 0.25mm 0.25 μm;
column temperature: the initial temperature is 100 ℃, the temperature is kept for 1min, the temperature is increased to 160 ℃ at the speed of 30 ℃/min, and the temperature is kept for 5 min; then heating to 310 ℃ at the speed of 30 ℃/min, and keeping for 17 min;
sample inlet temperature: the split ratio is 30 at 300 ℃;
detector temperature: 320 ℃;
carrier gas: high purity N2(purity 99.999%), flow rate 1.1 mL/min;
sample introduction amount: 1 μ L.
As shown in figure 2, the change of SCCPs before and after 14 days of degradation of SCCPs by Escherichia coli is shown, and the removal rate of SCCPs can reach 89.88% after 14 days of degradation of SCCPs by Escherichia coli is shown by the detection result; it is fully demonstrated that the strain with the specific sequence and composition can realize high-efficiency degradation effect of 89.88% in removal rate of SCCPs in a short time.
Ion chromatography conditions (ICS-600):
the type of the chromatographic column: anion chromatography column IonPacAS23, internal diameter 4X 250 mm;
leacheate: 4.5mM Na2CO3/0.8mM NaHCO3；
Flow rate: 1.0 mL/min;
column temperature: 30 ℃;
suppression of current: 25 mA;
carrier gas: high purity N2(purity 99.999%), flow rate 1.1 mL/min;
sample introduction amount: 10 μ L.
As shown in FIG. 3, it is known from the experimental results that the change of chloride ions before and after 14 days of SCCPs degradation by Escherichia coli, Cl removed from SCCPs into solution after 14 days of SCCPs degradation by Escherichia coli-The content is 11.71 mg/L; it is fully demonstrated that the specific sequences and compositions of the present invention provide strains with short time periods for the removal of Cl from SCCPs into solution-High-efficiency degradation effect with the content of 11.71 mg/L.
Sequence listing
<110> Zhejiang Ningbo theory of technology, college
<120> bacterial strain for effectively degrading short-chain chlorinated paraffin, and rapid screening method and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1472
<212> DNA
<213> Escherichia coli (Escherichia coli)
<400> 1
tcagattgaa cgctggcggc aggcctaaca catgcaagtc gaacggtaac aggaagaagc 60
ttgcttcttt gctgacgagt ggcggacggg tgagtaatgt ctgggaaact gcctgatgga 120
gggggataac tactggaaac ggtagctaat accgcataac gtcgcaagac caaagagggg 180
gaccttcggg cctcttgcca tcggatgtgc ccagatggga ttagctagta ggtggggtaa 240
cggctcacct aggcgacgat ccctagctgg tctgagagga tgaccagcca cactggaact 300
gagacacggt ccagactcct acgggaggca gcagtgggga atattgcaca atgggcgcaa 360
gcctgatgca gccatgccgc gtgtatgaag aaggccttcg ggttgtaaag tactttcagc 420
ggggaggaag ggagtaaagt taataccttt gctcattgac gttacccgca gaagaagcac 480
cggctaactc cgtgccagca gccgcggtaa tacggagggt gcaagcgtta atcggaatta 540
ctgggcgtaa agcgcacgca ggcggtttgt taagtcagat gtgaaatccc cgggctcaac 600
ctgggaactg catctgatac tggcaagctt gagtctcgta gaggggggta gaattccagg 660
tgtagcggtg aaatgcgtag agatctggag gaataccggt ggcgaaggcg gccccctgga 720
cgaagactga cgctcaggtg cgaaagcgtg gggagcaaac aggattagat accctggtag 780
tccacgccgt aaacgatgtc gacttggagg ttgtgccctt gaggcgtggc ttccggagct 840
aacgcgttaa gtcgaccgcc tggggagtac ggccgcaagg ttaaaactca aatgaattga 900
cgggggcccg cacaagcggt ggagcatgtg gtttaattcg atgcaacgcg aagaacctta 960
cctggtcttg acatccacag aactttccag agatggattg gtgccttcgg gaactgtgag 1020
acaggtgctg catggctgtc gtcagctcgt gttgtgaaat gttgggttaa gtcccgcaac 1080
gagcgcaacc cttatccttt gttgccagcg gtccggccgg gaactcaaag gagactgcca 1140
gtgataaact ggaggaaggt ggggatgacg tcaagtcatc atggccctta cgaccagggc 1200
tacacacgtg ctacaatggc gcatacaaag agaagcgacc tcgcgagagc aagcggacct 1260
cataaagtgc gtcgtagtcc ggattggagt ctgcaactcg actccatgaa gtcggaatcg 1320
ctagtaatcg tggatcagaa tgccacggtg aatacgttcc cgggccttgt acacaccgcc 1380
cgtcacacca tgggagtggg ttgcaaaaga agtaggtagc ttaaccttcg ggagggcgct 1440
taccactttg tgattcatga ctggggtgaa gt 1472
Claims (6)
1. A bacterial strain for effectively degrading short-chain chlorinated paraffin is characterized in that: the strain is SCCPs degrading bacteria, and is named in classification: escherichia coli (Escherichia coli) with a deposit number of: CGMCC No. 20989.
2. The strain for effectively degrading short-chain chlorinated paraffin according to claim 1, wherein: the gene sequence of the strain is specifically the gene sequence shown in SEQ ID NO. 1.
3. Use of a strain according to any of claims 1-2 for degrading short-chain chlorinated paraffins.
4. The method for screening the strain capable of effectively degrading the short-chain chlorinated paraffin according to any one of claims 1-2, wherein the strain comprises the following steps: the method comprises the following specific steps:
(1) adopting 1000mg/L sodium acetate as a carbon source of a culture medium, and simultaneously adding a nitrogen source, a phosphorus source and trace elements;
(2) filtering activated sludge, adding the filtrate into a pressurized dissolved oxygen reactor, adding a large amount of SCCPs with the concentration of 50-200mg/L, adjusting the pressure to 0.3MPa by oxygenation, and performing acclimation for 2-3d in the culture medium in the step (1); then carrying out plate streaking on a beef extract peptone solid culture medium, and selecting strains which can grow out on the culture medium;
(3) re-screening of strains: selecting strains capable of growing in a beef extract peptone solid medium, introducing the strains into a liquid medium, wherein the liquid medium is the same as the domestication medium, adding SCCPs with the concentration of 20mg/L, and performing an SCCPs degradation experiment.
5. The method for screening strains effective for degrading short-chain chlorinated paraffin according to claim 4, wherein: the specific concentrations of the nitrogen source, the phosphorus source and the trace elements added in the step (1) are as follows: 825mg/L (NH)4)2SO4，340mg/L KH2PO4，10mg/L MgSO4，20mg/L Fe2(SO4)3，4mg/L ZnSO4，3.2mg/L MnSO4，1mg/L CuSO4。
6. The method for screening strains effective for degrading short-chain chlorinated paraffin according to claim 4, wherein: the specific preparation process of the beef extract peptone solid medium in the step (2) comprises the following steps: adding SCCPs with the final concentration of 50mg/L into a beef extract peptone solid culture medium, then carrying out autoclaving at 121 ℃ for 30min, subpackaging the culture medium and pouring into a culture dish, and scratching a plate after the culture medium is solidified.
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