CN102952767B - Pseudomonas psychrophila HA-4 low-temperature sulfamethoxazole degrading bacteria as well as screening method and application thereof - Google Patents

Pseudomonas psychrophila HA-4 low-temperature sulfamethoxazole degrading bacteria as well as screening method and application thereof Download PDF

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CN102952767B
CN102952767B CN201210153552.7A CN201210153552A CN102952767B CN 102952767 B CN102952767 B CN 102952767B CN 201210153552 A CN201210153552 A CN 201210153552A CN 102952767 B CN102952767 B CN 102952767B
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sulfamethoxazole
strain
temperature
low
pseudomonas
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CN102952767A (en
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马放
蒋本超
李昂
蔡蕊
崔迪
邱天
张斯�
庞长泷
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Yixing Environmental Protection Industry Co.,Ltd.
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HIT YIXING ACADEMY OF ENVIRONMENTAL PROTECTION
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Abstract

The invention discloses a pseudomonas psychrophila HA-4 low-temperature sulfamethoxazole degrading bacteria, and belongs to the technical fields of bioengineering and environment engineering technology. The pseudomonas psychrophila HA-4 low-temperature sulfamethoxazole degrading bacteria is a low-temperature pseudomonas strain, which is preserved by the microbial strains preservation management committee center of general microbiology in March 27th, 2012, with the preservation number of CGMCC No.5944. The pseudomonas psychrophila HA-4 low-temperature sulfamethoxazole degrading bacteria is obtained by separating sludge in some sewage treatment plant aeration tank in Haerbin city; the strain can be grown by serving sulfamethoxazole as unique carbon, nitrogen and energy sources; the sulfamethoxazole can be degraded during culture of cell liquid; the cell liquid can be used for biodegradation of the sulfamethoxazole in city sewage; and by the strain, the removal rate of the sulfamethoxazole with the initial concentration of 100mg/L under a low-temperature condition is up to 35.34 percent.

Description

One strain Pseudomonas psychrophila HA-4 low temperature sulfamethoxazole degradation bacteria and screening method and application
Technical field
The invention belongs to biotechnology, field of environment engineering technology, relate to bacterial strain pseudomonas psychrophilahA-4 and the application in degraded sulfamethoxazole thereof.
Background technology
Medicine and personal-care supplies (Pharmaceuticals and personal care products) mainly comprise various prescribed and non prescribed medicines, spices, hair dye, makeup, opalizer, hair jelly, shampoo, perfumed soap etc.Wherein most of medicine is just directly drained in environment without metabolism in vivo, in addition in the production process of medicine and process and expiredly also can cause it to enter in water surrounding with not using in the process of medicine; Personal-care supplies also can directly enter water surrounding when washing one's face and rinsing one's mouth and swim.
Although the concentration that most PPCPs exist in environment is very low, due to bioconcentration, along with the increase of time, PPCPs will be finally to the ecosystem, especially water surrounding is produced to far-reaching and expendable impact.The treatment process that in water body, PPCPs is conventional has Physical, chemical method and biological process etc., and biological treatment is compared and is considered to environmental friendliness and method with low cost the most.The biological treatment of PPCPs with the concentration of the structure of its compound, starting point concentration, elementary substrate, the source of microbe inoculation and incubation time are relevant.
Sulfamethoxazole is widely used in human body as a kind of antibacterials, and it belongs to a kind of in PPCPs.At present, in China's effluent of municipal sewage plant, the mean concns level of this medicine is the highest, even has the negative phenomenon of removing to exist, and in the winter time will be higher than spring in its mean concns of northern area.
Foreign study shows, bacillus firmus, bacillus cereus, rhodococcus rhodochrous, pseudomonas, flavobacterium, aeromonasdeng the ability with degraded sulfamethoxazole.At present, both at home and abroad researcher mainly studies the biological degradation of sulfamethoxazole under normal temperature condition and report under cold condition is less.
Summary of the invention
Object of the present invention is exactly to remove and take the usefulness of the PPCPs pollutant that sulfamethoxazole is representative for strengthening low temp biochemical system, from occurring in nature, isolates bacterial strain pseudomonas psychrophilahA-4 can promote the degraded of sulfamethoxazole in low temperature environment, the removal based theoretical to the sulfamethoxazole in low temperature municipal effluent.It is a strain low temperature pseudomonas strain.
Technical solution of the present invention is: a strain pseudomonas psychrophilahA-4 low temperature sulfamethoxazole degradation bacteria, it is characterized in that a strain low temperature pseudomonas strain, is preserved in " China Committee for Culture Collection of Microorganisms's common micro-organisms " center ", its preserving number CGMCC No.5944 on March 27th, 2012.
Bacterial strain pseudomonas psychrophilahA-4 is that separation obtains from active sludge, and design temperature (10 ℃) screening is specifically screened step as follows:
(1) getting the 20mL Harbin fresh active sludge of peaceful municipal wastewater treatment plant Aerobic Pond adds 100mL to contain 20mg/L to take in the minimal medium triangular flask that sulfamethoxazole is sole carbon source, the energy, being placed in temperature is 10 ℃, rotating speed is to cultivate one week in the shaking table of 150r/min, obtains enrichment bacterium liquid;
(2) get 10mL enrichment bacterium liquid and add in 100mL sulfamethoxazole minimal medium triangular flask, the culture condition of repeating step (1) 3 ~ 4 times, the concentration of each sulfamethoxazole improves step by step, finally to 100mg/L, obtains domestication bacterium liquid;
(3) get 100 μ L domestication bacterium liquid and add and contain in 100mg/L sulfamethoxazole solid inorganic salt culture medium flat plate, coating method smoothens, and is placed in temperature and is the incubator of 10 ℃, cultivates one week, obtains single bacterium colony;
(4) repetitive operation step (3) 3 ~ 4 times, completes the acquisition of purebred low temperature sulfamethoxazole degradation bacteria;
(5) the purebred low temperature sulfamethoxazole degradation bacteria filtering out is inoculated into inclined-plane enrichment medium, in 4 ounder C, preserve inoculum size 10%;
Wherein the composition of minimal medium is: 3.5g K 2hPO 4, 0.5gNaCl, 1.5g KH 2pO 4, 0.15g MgSO 47H 2o, 0.5g (NH 4) 2sO 4, 1mL trace element solution is dissolved in 1000mL distilled water, regulates pH7.0; The composition of inorganic salt solid medium is in the composition of minimal medium, separately to add agar 2g/L;
Enrichment culture based component is: 1.5 g KH 2pO 4, 0.15 g MgSO 47H 2o, 0.5 g (NH 4) 2sO 4, 0.5 g NaCl, 3.5 g K 2hPO 4, 2g peptone, 18g agar, 1.0 mL trace element solutions be dissolved in the distilled water of 1000mL, regulates pH to 7.0;
The composition of trace element solution is: 0.1 g CuSO 45H 2o, 0.3 g MnSO 44H 2o, 0.2 g ZnSO 47H 2o, 2.0 g NaHCO 310H 2o, 0.5 g CoCl 26H 2o, 0.05g CaCl 22H 2o, 0.5g FeSO 47H 2o, 0.02g (NH 4) 6mo 7o 24H 2o is dissolved in the distilled water of 1000mL.
The OD of the domestication bacterium liquid that step (2) obtains 660=1.192; Substratum is all at 121 ℃, and autoclaving was used after 20 minutes.
This bacterial strain pseudomonas psychrophilahA-4 is suitable to grow in the developing medium of (pH 6.0), and growth temperature is 10 ℃, its bacteriology morphological specificity: be gram negative bacillus, atrichia, aerobic, degradable sulfamethoxazole, can take sulfamethoxazole as sole carbon source and energy growth.
Through strain identification, the 16S rDNA sequence that to have obtained length be 1609bp.
PCR primer adopts universal primer F8 (5 '-AGA GTT TGA TCA TGG CTC AG-3) and the R1522 (5 '-AAG GAC GTC ATC CAG CCG CA-3 ') of bacterial 16 S rDNA.With PCR instrument (GeneAmp, PCR System 9700), carry out amplified reaction.Reaction system cumulative volume is 25 μ l: ultrapure water 16.2 μ l, and 10 х Buffer 2.5 μ l, dNTP 2 μ l, F8 primer 1 μ l, R22 primer 1 μ l, ExTaq enzyme 0.3 μ l, total DNA does to get 2 μ l in reaction system after certain dilution.94 ℃ of denaturation 5 min, through 30 circulations, wherein, and 98 ℃ of sex change 20 s, 55 ℃ of annealing 40 s, 72 ℃ are extended 1.5 min, after 30 circulations, extend 7 min, 1% agarose gel electrophoresis, ultraviolet detection again at 72 ℃.By Blast program, carry out homology comparison, show that the homology of this bacterial strain and Pseudomonas psychrophila is up to 99%.
Bacterial strain of the present invention pseudomonas psychrophilahA-4 is applied to the biological treatment of sulfamethoxazole in sewage under low temperature.
Bacterial strain of the present invention pseudomonas psychrophilahA-4 need tame before application in the minimal medium with sulfamethoxazole, culture condition be bacterial strain at 10 ℃, pH 7.0, low-temperature aerobic is cultivated.This bacterial strain has higher degradation capability to sulfamethoxazole in low temperature habitat, and has substrate broad spectrum widely, in Table 1:
The substrate broad spectrum of table 1 bacterial strain HA-4
Substrate Result Substrate Result
Carbamzepine + Carbazole +
17 α-lynoral + Oestrone +
Trihydroxy-oestrin + Pentanoic +
Oil of mirbane - P-diaminodiphenyl +
Resorcinol + 1-amino-2-an aromatic plant metioned in ancient books phenol-4-sulfonic acid +
Ibuprofen BP/EP + Dimethylbenzene -
Two different sour toluene esters + 4-amino-2,2,6,6-tetramethylpiperidine +
Toluene - Benzene -
Note :+: well-grown;-: do not grow
The beneficial effect that the present invention reaches is: strain Pseudomonas psychrophila HA-4 provided by the invention has the ability of higher degraded sulfamethoxazole, under bacterial strain optimal growth condition, sulfamethoxazole to 100 mg/L, at short notice, degradation rate reaches more than 35.34%.
Accompanying drawing explanation
Fig. 1 is bacterial strain of the present invention pseudomonas psychrophilathe stereoscan photograph of HA-4.
Fig. 2 is bacterial strain of the present invention pseudomonas psychrophilahA-4 take sulfamethoxazole in unique carbon and nitrogen sources substratum, grow with sulfamethoxazole degradation curve figure.
Fig. 3 is the degradation curve figure of bacteria suspension of the present invention to different starting point concentration sulfamethoxazoles.
To be pseudomonas strain of the present invention take the determining of sulfamethoxazole that starting point concentration the is 100mg/L best pH growth conditions in unique carbon and nitrogen sources substratum to Fig. 4.
To be pseudomonas strain of the present invention take the determining of sulfamethoxazole that starting point concentration the is 100mg/L optimum temps growth conditions in unique carbon and nitrogen sources substratum to Fig. 5.
To be pseudomonas strain of the present invention take the determining of sulfamethoxazole that starting point concentration the is 100mg/L optimum revolution growth conditions in unique carbon and nitrogen sources substratum to Fig. 6.
To be pseudomonas strain of the present invention take the determining of sulfamethoxazole that starting point concentration the is 100mg/L optimum inoculation amount growth conditions in unique carbon and nitrogen sources substratum to Fig. 7.
Embodiment
Below in conjunction with accompanying drawing, the invention will be further described.
Embodiment 1
Strain Pseudomonas psychrophila provided by the invention separation from active sludge obtains, and design temperature (10 ℃) screening is specifically screened step as follows:
(1) getting the 20mL Harbin fresh active sludge of peaceful municipal wastewater treatment plant Aerobic Pond adds 100mL to contain 20mg/L to take in the minimal medium triangular flask that sulfamethoxazole is sole carbon source, the energy, being placed in temperature is 10 ℃, rotating speed is to cultivate one week in the shaking table of 150r/min, obtains enrichment bacterium liquid;
(2) getting 10mL enrichment bacterium liquid adds in 100mL sulfamethoxazole minimal medium triangular flask, the culture condition of repeating step (1) 3 ~ 4 times, the concentration of each sulfamethoxazole improves step by step, finally to 100mg/L, obtain domestication bacterium liquid, the OD of the domestication bacterium liquid of acquisition 660=1.192;
(3) get 100 μ L domestication bacterium liquid and add and contain in 100mg/L sulfamethoxazole solid inorganic salt culture medium flat plate, coating method smoothens, and is placed in temperature and is the incubator of 10 ℃, cultivates one week, obtains single bacterium colony;
(4) repetitive operation step (3) 3 ~ 4 times, completes the acquisition of purebred low temperature sulfamethoxazole degradation bacteria;
(5) the purebred low temperature sulfamethoxazole degradation bacteria filtering out is inoculated into inclined-plane enrichment medium, in 4 ounder C, preserve inoculum size 10%;
Wherein the composition of minimal medium is: 3.5g K 2hPO 4, 0.5gNaCl, 1.5g KH 2pO 4, 0.15g MgSO 47H 2o, 0.5g (NH 4) 2sO 4, 1mL trace element solution is dissolved in 1000mL distilled water, regulates pH7.0; The composition of inorganic salt solid medium is in the composition of minimal medium, separately to add agar 2g/L;
Enrichment culture based component is: 1.5 g KH 2pO 4, 0.15 g MgSO 47H 2o, 0.5 g (NH 4) 2sO 4, 0.5 g NaCl, 3.5 g K 2hPO 4, 2g peptone, 18g agar, 1.0 mL trace element solutions be dissolved in the distilled water of 1000mL, regulates pH to 7.0;
The composition of trace element solution is: 0.1 g CuSO 45H 2o, 0.3 g MnSO 44H 2o, 0.2 g ZnSO 47H 2o, 2.0 g NaHCO 310H 2o, 0.5 g CoCl 26H 2o, 0.05g CaCl 22H 2o, 0.5g FeSO 47H 2o, 0.02g (NH 4) 6mo 7o 24H 2o is dissolved in the distilled water of 1000mL.
Substratum is all at 121 ℃, and autoclaving was used after 20 minutes.
This bacterial strain pseudomonas psychrophilahA-4 is suitable to grow in the developing medium of (pH 6.0), and growth temperature is 10 ℃, its bacteriology morphological specificity: be gram negative bacillus, atrichia, aerobic, degradable sulfamethoxazole, can take sulfamethoxazole as sole carbon source and energy growth.
This bacterial strain pseudomonas psychrophilahA-4 cultural characteristic:
This bacterium take on the solid medium that sulfamethoxazole is sole carbon source 10 ℃ cultivate 6-7 days, bacterium colony is little, rounded, neat in edge, projection slightly, bacterium colony is creamy white, and very easily provokes.Scanning electron microscopic observation is tyrothricin (see figure 1).
Bacterial strain provided by the invention pseudomonas psychrophilathe 16S rDNA sequence of HA-4.
PCR primer adopts universal primer F8 (5 '-AGA GTT TGA TCA TGG CTC AG-3) and the R1522 (5 '-AAG GAC GTC ATC CAG CCG CA-3 ') of bacterial 16 S rDNA.With PCR instrument (GeneAmp, PCR System 9700), carry out amplified reaction.Reaction system cumulative volume is 25 μ l: ultrapure water 16.2 μ l, and 10 х Buffer 2.5 μ l, dNTP 2 μ l, F8 primer 1 μ l, R22 primer 1 μ l, ExTaq enzyme 0.3 μ l, total DNA does to get 2 μ l in reaction system after certain dilution.94 ℃ of denaturation 5 min, through 30 circulations, wherein, and 98 ℃ of sex change 20 s, 55 ℃ of annealing 40 s, 72 ℃ are extended 1.5 min, after 30 circulations, extend 7 min, 1% agarose gel electrophoresis, ultraviolet detection again at 72 ℃.By Blast program, carry out homology comparison, show this bacterial strain with pseudomonas psychrophilahomology up to 99%, sequence is as follows:
TTGCATGCCTGCAGGTCGACGATTAGAGTTTGATCCTGGCTCAGATTGAACGCTGGCGGCAGGCCTAACACATGCAAGTCGAGCGGTAGAGAGGTGCTTGCACCTCTTGAGAGCGGCGGACGGGTGAGTAATACCTAGGAATCTGCCTGATAGTGGGGGATAACGTTCGGAAACGGACGCTAATACCGCATACGTCCTACGGGAGAAAGCAGGGGACCTTCGGGCCTTGCGCTATCAGATGAGCCTAGGTCGGATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCTACGATCCGTAACTGGTCTGAGAGGATGATCAGTCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGAAAGCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGTCTTCGGATTGTAAAGCACTTTAAGTTGGGAGGAAGGGCATTAACCTAATACGTTAGTGTTTTGACGTTACCGACAGAATAAGCACCGGCTAACTCTGTGCCAGCAGCCGCGGTAATACAGAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCGCGTAGGTGGTTTGTTAAGTTGAATGTGAAATCCCCGGGCTCAACCTGGGAACTGCATCCAAAACTGGCAAGCTAGAGTATGGTAGAGGGTAGTGGAATTTCCTGTGTAGCGGTGAAATGCGTAGATATAGGAAGGAACACCAGTGGCGAAGGCGACTACCTGGACTGATACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCAACTAGCCGTTGGGAGTCTTGAACTCTTAGTGGCGCAGCTAACGCATTAAGTTGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGCCTTGACATCCAATGAACTTTCTAGAGATAGATTGGTGCCTTCGGGAACATTGAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGTAACGAGCGCAACCCTTGTCCTTAGTTACCAGCACGTAATGGTGGGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGGCCTGGGCTACACACGTGCTACAATGGTCGGTACAAAGGGTTGCCAAGCCGCGAGGTGGAGCTAATCCCATAAAACCGATCGTAGTCCGGATCGCAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTAATCGTGAATCAGAATGTCACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGCACCAGAAGTAGCTAGTCTAACCTTCGGGAGGACGGTTACCACGGTGTGATTCATGACTGGGGTGAAGTCGTAACAAGGTAGCCGTAGGGGAACCTGCGGCTGGATCACCTCCTTAATCTCTAGAGGATCCCCGGGTACCGAGCT
Embodiment 2
The present invention take sulfamethoxazole in unique carbon and nitrogen sources substratum to its degraded and bacterial strain pseudomonas psychrophilathe application of HA-4 upgrowth situation research, its step is as follows:
Bacterial strain pseudomonas psychrophilahA-4 is at 10 ℃ of temperature, initial pH value 6.0, shaking speed 150r/min, under the condition of 50mL Erlenmeyer flask liquid amount 20mL, the inoculum size of HA-4 bacteria suspension is 15%, the starting point concentration 100mg/L of sulfamethoxazole degraded system, cultured continuously 192h, every 24h detects the concentration of sulfamethoxazole.Adopt high performance liquid chromatography, chromatographic condition is to adopt Waters e2695 Alliance HPLC, and chromatographic column is Symmetry -C18 post (4.6 * 250 mm, 5 μ m); Moving phase is acetonitrile: formic acid water=40:60(V/V); Detect wavelength 265nm; 30 ℃ of column temperatures; Flow velocity 1.0mL/min; Sample size 10 μ L; Retention time 8min.The degradation rate that records sulfamethoxazole in 192h is 35.34%.
Preparation HA-4 bacteria suspension:
The HA-4 bacterium liquid of cultivating with enrichment medium is divided and installed in centrifuge tube, centrifugal 15 min under 8000 r/min conditions, abandoning supernatant, then in centrifuge tube, add sterilized phosphate buffer solution, stir evenly the thalline of centrifuge tube bottom settlings, recentrifuge, so washs thalline 3 times, to remove the substratum of thalline surface adsorption.The thalline obtaining is configured with minimal medium, obtain bacteria suspension.Fig. 2 is bacterial strain pseudomonas psychrophilathe growth of HA-4 in enrichment medium and the degraded situation of sulfamethoxazole; As seen from the figure, the degraded of sulfamethoxazole occurs in the logarithmic phase of strain growth.
Embodiment 3
Sulfamethoxazole application to its Study on degradation in unique carbon and nitrogen sources substratum is being take in the present invention:
By the starting point concentration of sulfamethoxazole degraded system in alternate embodiment 2, other step is with embodiment 2.Fig. 3 is the degraded situation of HA-4 bacteria suspension to different starting point concentration sulfamethoxazoles; As seen from Figure 3, bacterial strain pseudomonas psychrophilahA-4 has efficient degradation capability to sulfamethoxazole, can tolerate sulfamethoxazole concentration up to 200mg/L.
Embodiment 4
The application of the optimal growth condition research of pseudomonas strain of the present invention:
By take sulfamethoxazole in embodiment 2 as unique carbon and nitrogen sources substratum is changed to enrichment medium, change pH, temperature, shaking speed and inoculum size, other step is with embodiment 2.From Fig. 4, Fig. 5, Fig. 6 and Fig. 7, in the sulfamethoxazole degraded system of starting point concentration 100mg/L, cultured continuously 192h, bacterial strain pseudomonas psychrophilathe optimal growth condition of HA-4,10 ℃ of temperature, pH=6.0, shaking speed 150r/min, inoculum size are 10%.
Yixing environmental protection research institute of <110> Harbin Institute of Technology
<120> mono-strain pseudomonas psychrophilahA-4 low temperature sulfamethoxazole degradation bacteria and screening method and application
<140>?201210153552.7
<141>?2012-05-17
<160>?1583
<210>?1
<211>?1583
<212>?DNA
<213> Rhodopseudomonas ( pseudomonassp.)
<220>
<400>?1
ttgcatgcct?gcaggtcgac?gattagagtt?tgatcctggc?tcagattgaa?cgctggcggc?60
aggcctaaca?catgcaagtc?gagcggtaga?gaggtgcttg?cacctcttga?gagcggcgga?120
cgggtgagta?atacctagga?atctgcctga?tagtggggga?taacgttcgg?aaacggacgc?180
taataccgca?tacgtcctac?gggagaaagc?aggggacctt?cgggccttgc?gctatcagat?240
gagcctaggt?cggattagct?agttggtgag?gtaatggctc?accaaggcta?cgatccgtaa?300
ctggtctgag?aggatgatca?gtcacactgg?aactgagaca?cggtccagac?tcctacggga?360
ggcagcagtg?gggaatattg?gacaatgggc?gaaagcctga?tccagccatg?ccgcgtgtgt?420
gaagaaggtc?ttcggattgt?aaagcacttt?aagttgggag?gaagggcatt?aacctaatac?480
gttagtgttt?tgacgttacc?gacagaataa?gcaccggcta?actctgtgcc?agcagccgcg?540
gtaatacaga?gggtgcaagc?gttaatcgga?attactgggc?gtaaagcgcg?cgtaggtggt?600
ttgttaagtt?gaatgtgaaa?tccccgggct?caacctggga?actgcatcca?aaactggcaa?660
gctagagtat?ggtagagggt?agtggaattt?cctgtgtagc?ggtgaaatgc?gtagatatag?720
gaaggaacac?cagtggcgaa?ggcgactacc?tggactgata?ctgacactga?ggtgcgaaag?780
cgtggggagc?aaacaggatt?agataccctg?gtagtccacg?ccgtaaacga?tgtcaactag?840
ccgttgggag?tcttgaactc?ttagtggcgc?agctaacgca?ttaagttgac?cgcctgggga?900
gtacggccgc?aaggttaaaa?ctcaaatgaa?ttgacggggg?cccgcacaag?cggtggagca?960
tgtggtttaa?ttcgaagcaa?cgcgaagaac?cttaccaggc?cttgacatcc?aatgaacttt?1020
ctagagatag?attggtgcct?tcgggaacat?tgagacaggt?gctgcatggc?tgtcgtcagc?1080
tcgtgtcgtg?agatgttggg?ttaagtcccg?taacgagcgc?aacccttgtc?cttagttacc?1140
agcacgtaat?ggtgggcact?ctaaggagac?tgccggtgac?aaaccggagg?aaggtgggga?1200
tgacgtcaag?tcatcatggc?ccttacggcc?tgggctacac?acgtgctaca?atggtcggta?1260
caaagggttg?ccaagccgcg?aggtggagct?aatcccataa?aaccgatcgt?agtccggatc?1320
gcagtctgca?actcgactgc?gtgaagtcgg?aatcgctagt?aatcgtgaat?cagaatgtca?1380
cggtgaatac?gttcccgggc?cttgtacaca?ccgcccgtca?caccatggga?gtgggttgca?1440
ccagaagtag?ctagtctaac?cttcgggagg?acggttacca?cggtgtgatt?catgactggg?1500
gtgaagtcgt?aacaaggtag?ccgtagggga?acctgcggct?ggatcacctc?cttaatctct?1560
agaggatccc?cgggtaccga?gct?1583
 

Claims (3)

  1. One strain HA-4 low temperature sulfamethoxazole degradation bacteria ( pseudomonas psychrophila), it is characterized in that a strain low temperature pseudomonas strain, on March 27th, 2012, be preserved in " China Committee for Culture Collection of Microorganisms's common micro-organisms " center ", its preserving number CGMCC No.5944.
  2. 2. a strain HA-4 low temperature sulfamethoxazole degradation bacteria according to claim 1, it is characterized in that bacterial strain is suitable grows in the developing medium of pH 6.0, growth temperature is 10 ℃, its bacteriology morphological specificity: be gram negative bacillus, atrichia, aerobic, degradable sulfamethoxazole, can take sulfamethoxazole as sole carbon source and energy growth.
  3. 3. a strain HA-4 low temperature sulfamethoxazole degradation bacteria claimed in claim 1 is applied to the biological treatment of sulfamethoxazole in sewage under low temperature.
CN201210153552.7A 2012-05-17 2012-05-17 Pseudomonas psychrophila HA-4 low-temperature sulfamethoxazole degrading bacteria as well as screening method and application thereof Active CN102952767B (en)

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