CN105087425A - Halomonas sp. capable of degrading phenols and high-throughput screening method and application thereof - Google Patents
Halomonas sp. capable of degrading phenols and high-throughput screening method and application thereof Download PDFInfo
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Abstract
The invention discloses Halomonas sp. capable of degrading phenols and a high-throughput screening method and application thereof. The Halomonas sp. PH4-5 is preserved under CGMCC 10666. A bacterial liquid is cultured with a 48-pore deep-pore board, phenol concentration is measured by 96-pore microplate reader plate spectrophotometry based on 4-amino antipyrine colorimetry, moderately halophilic bacteria efficient in degrading phenols are screened from 43 environmental samples contaminated by high salinity and organics, the Halomonas sp. is separated and verified from an optimal flora E3, and a salt tolerant degrading mechanism of the Halomonas sp. is studied. Studying results show that quick screening of environmental degrading bacteria provides methodological basis and also provides technical support for the treatment of high-salinity phenolic wastewater.
Description
Technical field
The present invention relates to a kind of Halophilic Bacterium of degradation of phenol and screening method thereof and application.
Background technology
All containing a large amount of phenol in the waste water of the industry discharges such as oil coking, medicine, chemical industry, printing and dyeing, agricultural chemicals, Coal dressing.Phenol has high toxicity, must remove before therefore phenolic wastewater drains into receiving water body.Contain a large amount of inorganic salt again in this type of waste water simultaneously, add the difficulty of wastewater treatment, become a difficult problem of current phenolic wastewater treatment.
Biochemical method phenolic wastewater has certain effect, but high salt condition can make active sludge metabolic enzyme activity be obstructed, and causes organic removal rate to reduce.For the biological treatment of high salt phenolic wastewater, screen a kind of Phenol-degrading Bacteria Strains, be the key improving phenol biodegradation, utilize halophilic bacterium can effectively process high salt organic waste water.
Halophilic Bacterium is that a class is distributed widely in hypersaline environment, the extreme microorganism that can grow under 3% – 15%NaCl, there is certain degradation capability, many Halophilic Bacteriums have been isolated at present from hypersaline environment, but, conventional screen bacterium method dullness is loaded down with trivial details, cannot obtain the bacterial strain of high degradation capability, is difficult to meet the demands.
The normally dull and stereotyped coating of traditional sieve bacterium adds that repeated multiple times line is separated, or adopts shaking flask or test tube to screen, and the work of sieve bacterium is dull to be repeated, and is only limitted to screen several environmental sample.Conventional screen bacterium method adopts shaking flask or test tube to cultivate limited flora successively usually, then goes out single bacterium by dull and stereotyped coating and plate streak separation and purification.Such as, although more existing bibliographical information adopts conventional screen bacterium method to isolate addicted to salt or Facultative Halophiles from salt environment,
etal. (2007) are isolated a strain Facultative Halophiles Penicilliumchrysogenum and can be degraded the phenol of at least 300mg/L from Portuguese salt mine.Arulazhaganetal. (2010) have screened a salt-tolerant cultures from from the mixing salt water sample of India.HaddadiandShavandi (2013) is isolated a strain Halomonassp.strainPH2-2 from Iran the soil of petroleum pollution, under 7%NaCl, reach 95% to 400mg/L phenol clearance.But conventional screen bacterium method dullness is loaded down with trivial details, reagent needs consumption large, and sieve bacterium efficiency is low thus cause processing limited sample size.
Summary of the invention
The object of this invention is to provide a kind of Halophilic Bacterium of efficient degradation phenol and high-throughput screening method thereof and application, to overcome the defect that prior art exists, meet the needs of association area application.
Halophilic Bacterium Halomonassp.PH4-5 of the present invention is the bacterial classification that one belongs to Halomonas (Halomonas), it has the ability of degradation of phenol, this bacterial classification is separated from the mineralized waste of Shanghai Lao Gang refuse landfill, on March 26th, 2015 in the center preservation of China General Microbiological culture presevation management committee's common micro-organisms, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, preserving number is CGMCC10666.Classification And Nomenclature is Halomonas;
Bacterial classification of the present invention, has following characteristic:
Growth conditions on substratum:
The LB substratum improved: peptone 5g/L, yeast extract paste 2.5g/L; Sodium-chlor 100g/L; Agar powder 1.8g/L (30 DEG C, 72h);
Colony shape: circular;
Bacterium colony surface elevation: level and smooth, glossy;
Size: 0.5-1mm;
Tone: yellow.
Bacteria selection method of the present invention, comprises the steps:
(1) from by the environment of high salt and Organic pollutants, gather environmental sample, described environmental sample is soil sample or water sample;
In serum bottle, add environmental sample and substratum:
Contain in described substratum:
NaCl (100g/L), MgCl
2(0.5g/L), KH
2pO
4(0.45g/L), K
2hPO
4(0.9g/L), NH
4cl (0.3g/L), KCl (0.3g/L), phenol (100mg/L), yeast extract paste 0.2g/L and trace element (1g/L), pH is 7.0;
Table 1 trace element components
Serum bottle is placed in 30 DEG C, and under the condition of 150rpm, dark place is cultivated, and bottle top air, as oxygen sources, transfer after 3-4 time, the flora of acquisition sediment-free;
Switching: get the bacterium liquid after 5mL enrichment and proceed in new serum bottle, add the substratum that 45mL is fresh, inoculum size is 10%.
(2) NaCl concentration is respectively the deep hole that 3% (w/v), 5% (w/v), 8% (w/v), 10% (w/v), 12% (w/v) and 15% (w/v) are placed in 48 hole depth orifice plates, in every hole, containing phenol and MSM, phenol concentration is 500mg/L, MSM concentration is 1mL;
Prepare 6 48 hole depth orifice plates;
Be forwarded to by the culture of step (1) in 48 hole depth orifice plates, in 30 DEG C, 200rpm cultivates 72h;
(3) under above-mentioned same culture conditions, transfer after three times, adopt the 96 hole microplate reader plate spectrophotometries residue phenol concentration based on 4-AA colorimetric;
Phenol degrading characteristic relatively under different salinity, filters out bacterial classification of the present invention;
The present invention is separated acquisition bacterial classification, and following method can be adopted to identify:
(ABigen, Beijing, China) is extracted, 16SrDNA gene with test kit;
With primer 2 7F (5 '-AGAGTTTGATCCTGGCTCAG-3 ') and 1492R (5 '-GGTTACCTTGTTACGACTT-3 ') amplification.
50 μ L reaction systems contain: 2 × PCRTaqmix25 μ L, upstream primer (10 μm of ol/L) 1 μ L, downstream primer (10 μm of ol/l) 1 μ L, DNA profiling 1 μ L, stoning sour water 22 μ L.PCR adopts following reaction conditions: 94 DEG C of sex change 5min; 94 DEG C of sex change 1min, 54 DEG C of annealing 30s, 72 DEG C extend 1min, 30 circulations; Then 72 DEG C of 10min; Be cooled to 4 DEG C.PCR reaction is carried out at MastercyleGradientthermalcycler (Eppendorf, Shanghai, China).After PCR primer carries out purifying (DNApurificationkit, ABigen, Beijing, China), explanation (TaKaRa, Dalian, the China) fragment according to manufacturers is connected to pMD19-Tvector system.After connecting product conversion, E.coliDH5 α grows on Luria-Bertani (LB) substratum (containing 15g/L agar and 100 μ g/L ammonia benzyls), cultivates 12h in 37 DEG C.White dot is verified by primer M13-47 (5 '-CGCCAGGGTTTTCCCAGTCACGAC-3 ') and RV-M (5 '-GAGCGGATAACAATTTCACACAGG-3 ') PCR.The Insert Fragment of correct size carries out check order (BioSune, Shanghai, China).16SrRNA gene order BLAST (http://blast.ncbi.nlm.nih.gov/Blast.cgi) software analysis, sequence alignment and Phylogenetic Analysis Molecular Evolutionary Genetics Epidemiological Analysis software MEGA5.05 analyze.
(1) bacteria total DNA is extracted
Get sterile culture liquid 5mL, the centrifugal 2min of 12000rpm, with genome DNA extracting reagent kit (ABigen, Beijing, China) after abandoning supernatant, extract bacterial genomes STb gene.Test kit extracts genomic dna step:
1. add 480 μ L50mMEDTA to mix;
2. add 120 μ L N,O-Diacetylmuramidase mixings, incubation 60min (every 5min takes out and vibrates) in 37 DEG C of water-baths;
The centrifugal 2min of 3.12000rpm, after abandoning supernatant, add 600 μ L nucleus lysates, put upside down mixing, in 80 DEG C of water-baths, incubation 15min, is cooled to room temperature;
4. after adding 200 μ L albumen precipitations, high-speed and continuous vibration mixing 25s, ice bath 5min on turbula shaker, the more centrifugal 10min of 12000rpm, now more visible white precipitates at the bottom of pipe;
5. in the 2mL centrifuge tube after careful absorption supernatant to new sterilizing (inhaling a little) more as far as possible; Get in supernatant 600 μ L to 1.5mL conical centrifuge tube after recentrifuge 2min;
6. add 600 μ L room temperature Virahols, with mixing until there is thread white DNA pellet in gentle inversion 30 times, direct centrifugal 12000rpm2min, abandons supernatant;
7. add 0.5mL70% ethanol, put upside down and wash DNA several times, horizontally on thieving paper, blot residual ethanol after the centrifugal 1min of 12000rpm at once rapidly but softly fall supernatant; On clean bench, fan dries up, general 1h (can't smell alcohol smell);
8. add 40 μ LddH
2o, flicks tube wall mixing, places and get final product electrophoresis a little while.
(2) 16SrRNA gene amplification
With the bacteria total DNA of said extracted for template, carry out pcr amplification with 27F and 1492R total length primer.
27F：5′-AGAGTTTGATCCTGGCTCAG-3′；
1492R：5′-GGTTACCTTGTTACGACTT-3′；
The reaction system of pcr amplification and condition are in table 2, table 3.
Table 2PCR amplification reaction system
Table 3PCR reaction conditions
The pcr amplification product agarose gel electrophoresis inspection of 2%, after 110V28min, takes a picture with gel imaging system after the dyeing of Goldview staining fluid.
(3) purifying of pcr amplification product reclaims
Pcr amplification product sepharose reclaims test kit (ABigen, Beijing, China), to specifications step purifying.
1., under long-wave ultra violet lamp, with the scalpel of clean calcination on spirit lamp flame, the DNA agar block of required recovery is cut, put into 1.5mL centrifuge tube, weigh;
2. add the DD of 600 μ L;
10min is placed in 3.56 DEG C of water-baths, and every 2min whirlpool concussion is once accelerated agar block and dissolved;
4. solution adds in adsorption column EC, and room temperature places the centrifugal 45s of 1min, 12000rpm, then outwells the waste liquid in collection tube;
5. add 700 μ L rinsing liquid WB, the centrifugal 30s of 12000rpm, discards waste liquid; Add 500 μ L rinsing liquid WB again, the centrifugal 30s of 12000rpm, discards waste liquid;
6. adsorption column EC is put back in sky collection tube, the centrifugal 2min of 12000rpm;
7. take out adsorption column EC, put into clean centrifuge tube, add the elution buffer EB of 50 μ L in adsorption film middle part, room temperature places the centrifugal 1min of 2min, 12000rpm.
Reclaiming product can with the agarose gel electrophoresis inspection of 1%
(4) molecular cloning
1) the T/A clone of bacterial 16 S rRNAPCR product
The connection test kit that this experiment uses is purchased from TAKARA biotechnology company limited, and carrier is pMD-19T.
10 μ L ligation systems of bacterial 16 S rRNAPCR product are: PCR reclaims product 2 μ L, pMD-19Tvector1 μ L, ddH
2o2 μ L, ligase enzyme SolutionI5 μ L.
In advance ligase enzyme and carrier are placed in thawed on ice.Add said components successively in PCR pipe according to linked system, flick tube wall and make it mix, be of short durationly centrifugally placed on 4 DEG C of refrigerators and connect 16h.
2) product conversion is connected
The E.coliDH5 α competent cell that this experiment uses is from Shanghai Yi Teng company limited, and step of converting is as follows:
1. 100 μ L competent cells are placed in and add 10 μ L connecting fluids after 5min on ice, place 30min on ice;
Forward to rapidly in ice bath after 2.42 DEG C of water-bath heat shock 90s, place 2min, do not rock in attention process;
3. immediately in Bechtop, in every pipe, add 900 μ LLB substratum, 37 DEG C, 90min (LB substratum is placed on flame limit always) is cultivated in 200rpm concussion;
4., at aseptic Bechtop, when being cooled to Erlenmeyer flask non-scald on hand a little after LB substratum autoclaving, adding pre-configured ammonia benzyl solution, shake up, be down flat plate while hot; Draw competent cell triangular scraper that 100 μ L transform to be uniformly coated on flat board and to absorb completely to the bacterium liquid on flat board;
5. remain bacterium liquid centrifugal, abandon 800 μ L supernatants, remaining mixing, gets 70 μ L and is separately coated with flat board;
Slat chain conveyor 12h is inverted at 6.37 DEG C.
3) positive colony screening
The white dot getting grow on plates with the rifle choicest of sterilizing, drying, in the 50 μ L checking PCR reaction systems configured, carries out positive clone identification.Sequencing primer is:
M13-47：5′-CGCCAGGGTTTTCCCAGTCACGAC-3′；
RV-M：5′-GAGCGGATAACAATTTCACACAGG-3′；
The reaction system of checking pcr amplification and reaction conditions are in table 4, table 5.
Table 4PCR amplification reaction system
Table 5PCR reaction conditions
Detect amplified production with 1% agarose gel electrophoresis, imaging of taking pictures, and preserve analysis.Positive colony that picking inserts correct fragment send the platinum company that still checks order to check order.
(5) Phylogenetic Analysis
Sequencing result DNAman, BioEdit software is carried out editor to analyze, by similarity higher than 98% sequence be classified as the bacterial classification of same genus, sequence is compared by BLAST in GenBank database, gets 20 sequences that similarity is the highest, by MEGA5.05 analytical sequence and phylogenetic tree construction.
By above-mentioned authentication method, it is a kind of bacterial classification belonging to Halomonas that the present invention obtains bacterial classification, but with the bacterial classification of existing Halomonas, have remarkable difference, its difference is: source place is different.The present invention obtains the mineralized waste that bacterial classification derives from refuse landfill, and has excellent phenol degrading ability.
Screening method of the present invention, the method of the halophilic bacterium group of degradation of phenol is cultivated for a kind of high-throughput, bacterium salt tolerant and degradation capability are designed high flux screening flow process as screening pressure, rapid screening can be carried out from environmental sample, there is the halophilic bacterium group of optimum degradation capability under different salinity, greatly reduce shaking table and take volume and reagent needs consumption, thus achieve microminiaturized sieve bacterium, may be used for the degradation bacteria in the different sample of extensive rapid screening.
Bacterial classification of the present invention, has the excellent ability of degradation of phenol, can as the bacterial classification of the high salt phenolic wastewater of biological treatment;
By bacterial classification of the present invention, the method for microorganism of degradation of phenol, comprises the steps:
(1) first cultivate described bacterial classification, cultural method is as follows:
Bacterial strain is inoculated in the MSM liquid nutrient medium (MgCl of the improvement containing 15mL according to the inoculum size of 5%
2: 0.5g/L, KH
2pO
4: 0.45g/L, K
2hPO
4: 0.9g/L, NH
4cl:0.3g/L, KCl:0.3g/L, yeast extract paste: 5g/L, sodium-chlor is respectively 3%, 5%, 8%, 10%, 12% (w/v)) test tube in, often prop up cuvette cartridge liquid measure 5mL, 30 DEG C, 150rpm is cultured to lag phase;
Culture is all transferred in 50mL centrifuge tube, then 4000rpm, 20 DEG C of centrifugal 15min collect thalline, with aseptic isotonic solution, (inorganic ion composition is identical with used medium composition, do not add organic substance, 121 DEG C of autoclaving 20min in advance) wash thalline, then the same terms collected by centrifugation thalline, be resuspended in 10mL isotonic solution;
(2) transfer above-mentioned bacterium liquid to 50mLMSM, and add 500mg/L phenol as in the substratum of sole carbon source and energy substance, sodium chloride concentration is 3% – 12% (w/v), 30 DEG C, process 68h under 150rpm, can remove phenol, clearance reaches more than 94%.
The present invention adopts 48 hole depth orifice plates to cultivate bacterium liquid, based on 96 hole microplate reader plate benzene by spectrophotometry phenol concentration of 4-AA colorimetric, and first Application the method is from being screened the Halophilic Bacterium of efficient degradation phenol 43 environmental samples of high salt and Organic pollutants, be separated from optimum flora E3 and identify a strain Halophilic Bacterium, and studying its salt tolerant degradation mechanism.Result of study is that the rapid screening of environment degradable bacterium provides methodology foundation, also provides technical support for processing high salt phenolic wastewater simultaneously.
Accompanying drawing explanation
Fig. 1 is the 96 hole microplate reader plate spectrophotometry typical curves based on 4-AA colorimetric.
Fig. 2 is the degradation rate of different halophilic bacterium group Pyrogentisinic Acid.
Fig. 3 is the degradation curve of Halomonassp.PH4-5 Pyrogentisinic Acid under different salinity.
Embodiment
Embodiment 1
43 environmental samples (table 6) are acquired by the environment of high salt and Organic pollutants from 8 kinds.First be enriched in 125-mL serum bottle by screening method step (1), transfer and obtain the flora of sediment-free for 3 times afterwards.
Table 6 sample collecting information
43 cultures (No:A1 – H1) are switched in 48 hole depth orifice plates by screening method step (2), and in 30 DEG C, 200rpm cultivates 72h.Adopt according to screening method step (3) and remain phenol concentration based in 96 hole microplate reader plate spectrophotometry deep-well plates of 4-AA colorimetric.200 μ L bacterium liquid in 48 hole depth orifice plates are transferred in 96 hole microplate reader plates, the centrifugal 20min of monoblock 96 hole microplate reader plate 2500 × g, supernatant liquor is transferred in 96 new hole microplate reader plates, be diluted to 300 μ L by a certain percentage, add 3 μ L ammoniacal liquor damping fluids, add 4 – aminoantipyrene 6 μ L, mixing, add the 6 μ L Tripotassium iron hexacyanides again, fully place 10min after mixing, in 510nm wavelength place, microplate reader reads absorbance in each micropore.
Typical curve formula based on 96 hole microplate reader plate spectrophotometrys of 4-AA colorimetric is y=0.08808x+0.00203, R
2=0.9988 (see Fig. 1), the selection result display (see Fig. 2), in Fig. 2, X-coordinate is flora, and wherein, A1 ~ A6, B1 are the cultivation floras from Cha Er Han Salt Lake; B2 ~ B6, C1, C2 are the cultivation floras from Salt Lake in Xinjiang; C3 ~ C6, D1, D2 are the cultivation floras from grand celebration saline-alkali soil; D3 ~ D6, E1 ~ E6, F1, F2 is the cultivation flora from Shanghai Lao Gang refuse landfill; F3 ~ F6, G1 ~ G3 is the cultivation flora from beach, the East Sea; G4 ~ G6, H1 are the cultivation floras from Shanghai Gaoqiao petrochemical plant.
The phenol (initial phenol concentration is 500mg/L) that in 43 halophilic bacterium groups, 10 floras can be degraded more than 50% under 3%-10% (w/v) NaCl: numbering is followed successively by C3, C4, D6, E3, E4, E5, E6, F1, G4 and G5, they are from grand celebration saline-alkali soil, Shanghai Laogang Landfill and Shanghai Gaoqiao petrochemical plant (see table 7).
Table 7 sieves bacterium result
In these halophilic bacterium group, E3 degradation effect is best, 3%, and 5%, 8%, 10% and 12%NaCl (w/v) condition under, initial 500mg/L phenol clearance is respectively 96.0%, 99.2%, 99.1%, 95.4% and 88.5%, E3 from Shanghai Lao Gang refuse landfill mineralized waste, 1994 landfill time.Then, from optimum flora E3, isolating 5 strain list bacterium, the degradation characteristic of more different single bacterium Pyrogentisinic Acid by being repeatedly separated line, obtaining Halomonassp.PH4-5 bacterial strain and there is best phenol degrading ability in wide in range salinity range.
Embodiment 2
Halomonassp.PH4-5 is seeded in the MSM substratum containing 500mg/L phenol and 3%-12% (w/v) sodium-chlor, investigates the degraded of Pyrogentisinic Acid.
Halomonassp.PH4-5 optimum growh and degraded salinity are 5%.After this bacterial strain grows 68h in the MSM substratum containing 500mg/L phenol, 3%, during 5%, 8%, 10%, 12% (w/v) NaCl, phenol clearance is respectively 96.2%, 99.8%, 98.9%, 94.7% and 86.3% (see Fig. 3).This bacterial strain all reaches more than 94% to the phenol clearance of 500mg/L in 3% – 10% salinity range, can in wide in range salinity range the phenol of degrading high concentration, embody the advantage of Halophilic Bacterium, can be used for the biological treatment of high salt phenolic wastewater.
Claims (5)
1. Halophilic Bacterium (Halomonassp.) PH4-5 of degradation of phenol, preserving number is CGMCC10666.
2. the screening method of Halophilic Bacterium (Halomonassp.) PH4-5 of degradation of phenol according to claim 1, comprises the steps:
(1) from by the environment of high salt and Organic pollutants, environmental sample is gathered;
In serum bottle, add environmental sample and substratum:
Contain in described substratum:
NaCl (100g/L), MgCl
2(0.5g/L), KH
2pO
4(0.45g/L), K
2hPO
4(0.9g/L), NH
4cl (0.3g/L), KCl (0.3g/L), phenol (100mg/L), yeast extract paste 0.2g/L and trace element (1g/L), pH is 7.0;
Serum bottle is placed in 30 DEG C, and under the condition of 150rpm, dark place is cultivated, and bottle top air, as oxygen sources, transfer after 3-4 time, the flora of acquisition sediment-free;
Switching: get the bacterium liquid after 5mL enrichment and proceed in new serum bottle, add the substratum that 45mL is fresh, inoculum size is 10%.
(2) NaCl concentration is respectively the deep hole that 3% (w/v), 5% (w/v), 8% (w/v), 10% (w/v), 12% (w/v) and 15% (w/v) are placed in 48 hole depth orifice plates, in every hole, containing phenol and MSM, phenol concentration is 500mgL
-1, MSM concentration is 1mL;
Prepare 6 48 hole depth orifice plates;
Be forwarded to by the culture of step (1) in 48 hole depth orifice plates, in 30 DEG C, 200rpm cultivates 72h;
(3) under above-mentioned same culture conditions, transfer after three times, adopt the 96 hole microplate reader plate spectrophotometries residue phenol concentration based on 4-AA colorimetric;
Phenol degrading characteristic relatively under different salinity, can obtain described bacterial classification.
3. the application of Halophilic Bacterium (Halomonassp.) PH4-5 of degradation of phenol according to claim 1, is characterized in that, for high salt phenolic wastewater of carrying out a biological disposal upon.
4. application according to claim 3, is characterized in that, application method comprises the steps:
(1) first described bacterial classification is cultivated;
(2) transfer above-mentioned bacterium liquid to 50mLMSM, and add 500mg/L phenol as in the substratum of sole carbon source and energy substance, sodium chloride concentration is 3% ~ 12% (w/v), 30 DEG C, processes 68h under 150rpm, can remove phenol.
5. method according to claim 4, is characterized in that, cultural method is as follows:
Bacterial strain is inoculated in the MSM liquid nutrient medium (MgCl of the improvement containing 15mL according to the inoculum size of 5%
2: 0.5g/L, KH
2pO
4: 0.45g/L, K
2hPO
4: 0.9g/L, NH
4cl:0.3g/L, KCl:0.3g/L, yeast extract paste: 5g/L, sodium-chlor is respectively 3%, 5%, 8%, 10%, 12% (w/v)) test tube in, often prop up cuvette cartridge liquid measure 5mL, 30 DEG C, 150rpm is cultured to lag phase;
Culture is all transferred in 50mL centrifuge tube, then 4000rpm, 20 DEG C of centrifugal 15min collect thalline, with aseptic isotonic solution, (inorganic ion composition is identical with used medium composition, do not add organic substance, 121 DEG C of autoclaving 20min in advance) wash thalline, then the same terms collected by centrifugation thalline, is resuspended in 10mL isotonic solution.
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| CN112899192A (en) * | 2021-02-04 | 2021-06-04 | 华东理工大学 | BTEX degrading bacterium and screening method and application thereof |
| CN114538695A (en) * | 2020-11-24 | 2022-05-27 | 山东福洋生物科技股份有限公司 | Method for treating modified starch industrial wastewater by using microorganisms |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107164277A (en) * | 2016-06-27 | 2017-09-15 | 内蒙古大学 | A kind of Halomonas of degradation of phenol |
| CN107325981A (en) * | 2016-06-27 | 2017-11-07 | 内蒙古大学 | A kind of method of quick screening stability and high efficiency phenol degrading Halophiles |
| CN107325981B (en) * | 2016-06-27 | 2020-07-17 | 内蒙古大学 | Method for rapidly screening stable and efficient phenol degradation halophilic bacteria |
| CN107164277B (en) * | 2016-06-27 | 2020-08-25 | 内蒙古大学 | Halomonas for degrading phenol |
| CN109439602A (en) * | 2018-12-29 | 2019-03-08 | 中蓝连海设计研究院有限公司 | Halophilic vibrio YL5-2 and its microbial inoculum are degraded the application in conversion pollutant under high salt conditions |
| CN109534518A (en) * | 2018-12-29 | 2019-03-29 | 中蓝连海设计研究院有限公司 | A kind of high-salt wastewater biology membrane treatment process using Halophiles YL5-2 |
| CN109534518B (en) * | 2018-12-29 | 2021-09-14 | 中蓝连海设计研究院有限公司 | High-salinity wastewater biofilm treatment process using halophilic bacteria YL5-2 |
| CN109439602B (en) * | 2018-12-29 | 2021-09-14 | 中蓝连海设计研究院有限公司 | Vibrio halophilus YL5-2 and application of microbial inoculum thereof in degrading and converting pollutants under high-salt condition |
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| CN114538695B (en) * | 2020-11-24 | 2024-05-24 | 山东福洋生物科技股份有限公司 | Method for treating modified starch industrial wastewater by utilizing microorganisms |
| CN112899192A (en) * | 2021-02-04 | 2021-06-04 | 华东理工大学 | BTEX degrading bacterium and screening method and application thereof |
| CN112899192B (en) * | 2021-02-04 | 2022-09-09 | 华东理工大学 | BTEX degrading bacterium and screening method and application thereof |
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