CN101250491A - Streptomycesalbidoflavus having new metabolic characteristics and its use in biological degradation - Google Patents
Streptomycesalbidoflavus having new metabolic characteristics and its use in biological degradation Download PDFInfo
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- CN101250491A CN101250491A CN 200710157811 CN200710157811A CN101250491A CN 101250491 A CN101250491 A CN 101250491A CN 200710157811 CN200710157811 CN 200710157811 CN 200710157811 A CN200710157811 A CN 200710157811A CN 101250491 A CN101250491 A CN 101250491A
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- streptomycesalbidoflhaving
- mirbane
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- nitrobenzene
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
Abstract
The invention relates to streptomyces albidoflauvs with the metabolism characteristic and the application of the streptomyces albidoflauvs in biodegradation, which belongs to the biological engineering technical field. The invention separates and screens a strain of streptomyces albidoflauvs with the metabolism characteristic. The application of the invention has the advantages that the streptomyces albidoflauvs degrades nitrobenzene through a part of reduction ways and mineralizes by-products of pyridinecarboxylic acid in the degrading process, thereby complete degradation of the nitrobenzene is realized, which solves the bottleneck in reducing and degrading the nitrobenzene partially, the streptomyces albidoflauvs can effectively degrade nitrobenzene under the condition that the salinity is high and a plurality of organic matters are co-existed, the streptomyces albidoflauvs can take pyridinecarboxylic acid as an unique carbon, azote, and energy source to grow, the pyridinecarboxylic acid is finally mineralized into innocuous carbon dioxide and water, and the streptomyces albidoflauvs can be taken as biological intesified preparation and applied in processing nitrobenzene wastewater and pyridinecarboxylic acid wastewater with high salinity and complex components biologically.
Description
Technical field
The invention belongs to technical field of bioengineering, relate to Streptomycesalbidoflhaving and the application in oil of mirbane aerobic biodegradation and pyridine carboxylic acid aerobic biodegradation thereof that a strain has new metabolic characteristics.
Background technology
Oil of mirbane is widely used in the production of medicine, dyestuff, plastics, explosive etc., and the widespread use of oil of mirbane causes the nitrobenzene contamination environment.Nitrobenzene waste water can be handled by aerobe method.At present, the Microbial resources of aerobic degradation oil of mirbane have bacterium and fungi, and bacterium has pseudomonas, comamonas, bacillus, coryneform bacteria, staphylococcus, suis.
Oil of mirbane most important purposes in industry is to produce azoic dyestuff, and azo dye wastewater has characteristics such as salinity height, complicated component.High salinity is meant that the total dissolved solid thing (sodium-chlor) in the waste water is at least 3.5% (W/V); Complicated component is meant in the waste water that except that containing oil of mirbane, main organic pollutant also has aniline and phenol.When high salinity condition or the coexistence of multiple organic substance, above-mentioned nitrobenzene degradation bacterium can not grow, and therefore can not realize the degraded of oil of mirbane.
The nitrobenzene degradation bacterium of being reported at present passes through two kinds of approach degrading nitrobenzenes under aerobic condition: oxidative pathway or partial reduction approach, wherein partial reduction approach is more extensive.In isolating 155 strain bacterium such as Spain, 154 strains are by partial reduction approach degrading nitrobenzene, and 1 strain is by the oxidative pathway degrading nitrobenzene.This is because the sucting electronic effect of nitro causes cloud density decline on the phenyl ring, thereby has hindered the electric attack of oxidasic parent; Simultaneously, the nitro electronegativity strengthens, is easy to be reduced.Self structure just because of oil of mirbane causes present institute strain separated nearly all by partial reduction approach degrading nitrobenzene.Yet in the partial reduction degraded of oil of mirbane, pyridine carboxylic acid generates as by product, and pyridine carboxylic acid has bio-toxicity, can not be utilized by the nitrobenzene degradation bacterium, and the generation of pyridine carboxylic acid has hindered the aerobic degradation of oil of mirbane.
Pyridine carboxylic acid is a kind of N heterocyclization and thing, is mainly used in the production of medicine, agricultural chemicals, dyestuff etc., and the application of pyridine carboxylic acid causes the pyridine carboxylic acid contaminate environment.At present, the Microbial resources of aerobic degradation pyridine carboxylic acid have Arthrobacter and mycobacterium.
Summary of the invention
Purpose of the present invention is exactly this problem of by product pyridine carboxylic acid that can not degrade to occur at characteristics such as actual oil of mirbane trade effluent salinity height, complicated component and in the nitrobenzene degradation process, tame, filter out the oil of mirbane efficient degrading bacteria by scientific approach, in order to the nitrobenzene waste water of processing salinity height, complicated component, and the bottleneck in the solution nitrobenzene degradation process.Simultaneously, also provide efficient, practical Microbial resources for the aerobic biodegradation of pyridine carboxylic acid.
Technical solution of the present invention is, Streptomycesalbidoflhaving provided by the invention is preserved in " China Committee for Culture Collection of Microorganisms's common micro-organisms " center "; its preserving number CGMCC No.1759, classification called after Streptomyces albidoflavus on July 18th, 2006.Address: No. 13, North No.1 Row, Zhongguancun, Haidian District, Beijing City, postcode: 100080.
Described Streptomycesalbidoflhaving, its screening step is as follows: get the 50ml active sludge from Dalian Dye Factory aeration tank, the trisodium phosphate of adding 0.2% after vibration 5-10min smashes the active sludge flco on the vibrator, is got the 5ml active sludge as separating flocculation agent, adding contains shaking in the bottle of 95ml minimal medium, add oil of mirbane again as sole carbon, nitrogen, the energy, wherein the concentration of oil of mirbane is 200mg/L, carries out shaking table and cultivates, 30 ℃ of temperature, rotating speed 180r/min; When bacterium liquid becomes muddy, transfer, obtaining with oil of mirbane is the stable bacterium liquid of sole carbon, nitrogen, the energy; To stablize bacterium liquid dilution, on the solid medium that with oil of mirbane is sole carbon, nitrogen, the energy, be coated with, this solid medium will be put into illumination box, 30 ℃, cultivated 4-5 days, choose single bacterium colony; Isolate a strain and be with oil of mirbane sole carbon, nitrogen, the energy Streptomycesalbidoflhaving.
Described Streptomycesalbidoflhaving is characterized in that, this strain growth appropriate pH value is 6.0-9.0, and suitable growth temperature is 20-40 ℃; The bacterial strain fibrillae of spores is short, straight, gentle bent to volution, the spore Long Circle.
The application of described Streptomycesalbidoflhaving in the oil of mirbane aerobic biodegradation, it is characterized in that, Streptomycesalbidoflhaving immobilized cell, cell liquid culture are all by partial reduction approach degrading nitrobenzene, and the by product pyridine carboxylic acid in the mineralizing and degrading process, realize the thorough degraded of oil of mirbane, therefore can handle as biological reinforced preparation p-nitrophenyl waste water.
The application of described Streptomycesalbidoflhaving in the oil of mirbane aerobic biodegradation, it is characterized in that Streptomycesalbidoflhaving immobilized cell, cell liquid culture are all under the condition of NaCl mass concentration 0-50mg/L, effectively therefore degrading nitrobenzene can be handled the nitrobenzene waste water of high salinity as biological reinforced preparation.
The application of described Streptomycesalbidoflhaving in the oil of mirbane aerobic biodegradation, it is characterized in that, Streptomycesalbidoflhaving immobilized cell, cell liquid culture are all in aniline and oil of mirbane coexistence, under the condition of aniline mass concentration 0-50mg/L, effectively therefore degrading nitrobenzene can be handled the nitrobenzene waste water of complicated component as biological reinforced preparation.
The application of described Streptomycesalbidoflhaving in the oil of mirbane aerobic biodegradation, it is characterized in that, Streptomycesalbidoflhaving immobilized cell, cell liquid culture are all in phenol and oil of mirbane coexistence, under the condition of phenol mass concentration 0-200mg/L, effectively therefore degrading nitrobenzene can be handled the nitrobenzene waste water of complicated component as biological reinforced preparation.
The application of described Streptomycesalbidoflhaving in the pyridine carboxylic acid aerobic biodegradation, it is characterized in that, Streptomycesalbidoflhaving immobilized cell, cell liquid culture all destroy the N heterocycle of pyridine carboxylic acid, therefore pyridine carboxylic acid open loop mineralising can be handled pyridine carboxylic acid waste water as biological reinforced preparation.
Described Streptomycesalbidoflhaving physiological and biochemical property sees Table 1 and 2
The cultural characteristic of table 1 Streptomycesalbidoflhaving
| Cultural characteristic | Aerial hyphae | Substrate mycelium | Soluble pigment |
| Glucose asparagine substratum glycerine asparagine substratum inorganic salt starch culture-medium yeast powder malt meal substratum oatmeal substratum Gause I substratum mulberry Ta Shi substratum | The yellow dirt dust ash of the yellow shallow mustard of the white hawksbill turtle of lotus seeds junket coptis root is white | The yellow shallow pine soot bamboo shoot skin palm fibre bamboo shoot skin palm fibre freshwater mussel meat sinapsis alba of the yellow pomegranate calyx of shallow mustard is yellow withered green | There is not the no shark cyan of no cloves brown clove palm fibre |
The physio-biochemical characteristics of table 2 Streptomycesalbidoflhaving
| Test subject | The result | Test subject | The result |
| Utilization of carbon source: D-semi-lactosi synanthrin wood sugar cellobiose N.F,USP MANNITOL raffinose fructose sucrose sorbose lactose melibiose melizitose ribose | + - + + + - + - - + + - + | Utilization of carbon source (continuing): sorbyl alcohol glycerine pectinose seminose maltose trehalose starch gelatin hydrolysate liquefaction milk peptonizes old propylhomoserin enzyme nitrate reduction and produces hydrogen sulfide | - + + + + + + - + - + - |
Annotate :+: the positive;-: feminine gender
Described Streptomycesalbidoflhaving both can be grown in eutrophy substratum LB, also can grow in small organic molecules such as glucose, sucrose, starch are the minimal medium of sole carbon source.
Described Streptomycesalbidoflhaving can be with the bacterial classification or immobilized cell p-nitrophenyl waste water of making and pyridine carboxylic acid waste water biodegrade in vegetative period of fresh culture.
The beneficial effect that the present invention reached is: Streptomycesalbidoflhaving provided by the invention is by partial reduction approach degrading nitrobenzene, and the by product pyridine carboxylic acid in the mineralizing and degrading process, thereby realize the thorough degraded of oil of mirbane, solved the bottleneck in the oil of mirbane partial reduction degraded; Streptomycesalbidoflhaving is thorough degrading nitrobenzene under the condition of sole carbon, nitrogen, the energy and oil of mirbane starting point concentration≤400mg/L at oil of mirbane; Streptomycesalbidoflhaving is at NaCl mass concentration 0-50mg/L, and oil of mirbane is effective degrading nitrobenzene under the condition of sole carbon, nitrogen, the energy and oil of mirbane starting point concentration≤400mg/L; Streptomycesalbidoflhaving is in oil of mirbane and aniline coexistence, and aniline concentration is 0-50mg/L, under the condition of oil of mirbane starting point concentration≤400mg/L, and effective degrading nitrobenzene; Streptomycesalbidoflhaving is in oil of mirbane and phenol coexistence, and phenol concentration is 0-200mg/L, under the condition of oil of mirbane starting point concentration≤400mg/L, and effective degrading nitrobenzene; Streptomycesalbidoflhaving is thorough degraded pyridine carboxylic acid under the condition of sole carbon, nitrogen, the energy and pyridine carboxylic acid starting point concentration≤3000mg/L at pyridine carboxylic acid; Streptomycesalbidoflhaving provided by the invention can be used as biological bacteria preparation, is added in existing nitrobenzene waste water and the pyridine carboxylic acid Waste Water Treatment, strengthens the processing power of former processing system; Streptomycesalbidoflhaving provided by the invention has wide application potential in the processing of nitrobenzene waste water and pyridine carboxylic acid waste water.
Description of drawings
Fig. 1 is a Streptomycesalbidoflhaving provided by the invention is being that sole carbon, nitrogen, the energy and oil of mirbane starting point concentration are growth and nitrobenzene degradation graphic representation in the liquid culture of 400mg/L with oil of mirbane, the left side ordinate zou is represented the rate of recovery of oil of mirbane among the figure, unit is %, the right side ordinate zou is represented dry cell weight, and unit is mg/L;
Among the figure :-■-the represent rate of recovery ,--represent thalli growth amount.
Fig. 2 is a Streptomycesalbidoflhaving provided by the invention is being that sole carbon, nitrogen, the energy and pyridine carboxylic acid starting point concentration are growth and pyridine carboxylic acid degradation curve figure in the liquid culture of 3000mg/L with the pyridine carboxylic acid, the left side ordinate zou is represented the rate of recovery of pyridine carboxylic acid among the figure, unit is %, the right side ordinate zou is represented the increment of thalline, and unit is OD
660
Among the figure :-■-the represent rate of recovery ,--represent thalli growth amount.
Fig. 3 is that Streptomycesalbidoflhaving provided by the invention is dense down in different sodium-chlor quality, oil of mirbane is that sole carbon, nitrogen, the energy and oil of mirbane starting point concentration are when being 400mg/L, the degradation curve figure of oil of mirbane, ordinate zou is represented the rate of recovery of oil of mirbane among the figure, unit is %;
Among the figure :-■-represent sodium-chlor mass concentration 20mg/L ,--represent sodium-chlor mass concentration 30mg/L ,-zero-represent sodium-chlor mass concentration 50mg/L ,-●-represent sodium-chlor mass concentration 70mg/L.
Fig. 4 is a Streptomycesalbidoflhaving provided by the invention in aniline and oil of mirbane coexistence, and the oil of mirbane starting point concentration is when being 400mg/L, the degradation curve figure of oil of mirbane, and ordinate zou is represented the rate of recovery of oil of mirbane among the figure, unit is %;
Among the figure :-■-represent aniline concentration 25mg/L ,--represent aniline concentration 50mg/L ,-zero-represent aniline concentration 75mg/L ,-●-represent aniline concentration 100mg/L.
Fig. 5 is a Streptomycesalbidoflhaving provided by the invention in phenol and oil of mirbane coexistence, and the oil of mirbane starting point concentration is when being 400mg/L, the degradation curve figure of oil of mirbane, and ordinate zou is represented the rate of recovery of oil of mirbane among the figure, unit is %;
Among the figure :-■-represent phenol concentration 50mg/L ,--represent phenol concentration 100mg/L ,-zero-represent phenol concentration 150mg/L ,-●-represent phenol concentration 200mg/L.
Among Fig. 1 to 5: X-coordinate h all represents the time, and unit is hour.
Embodiment
The invention will be further described below in conjunction with accompanying drawing.
Embodiment 1
Streptomycesalbidoflhaving provided by the invention, its screening step is as follows:
From the aeration tank of Dalian Dye Factory, gather the aerobic activated sludge sample, tame, cultivate as the bacterium source.With the 50ml active sludge, the trisodium phosphate of adding 0.2% is as separating flocculation agent, after vibration 5-10min smashes the active sludge flco on the vibrator, get the 5ml active sludge, adding contains shaking in the bottle of 95ml minimal medium, adds oil of mirbane again as sole carbon, nitrogen, the energy, and wherein the concentration of oil of mirbane is 200mg/L, carry out shaking table and cultivate 30 ℃ of temperature, rotating speed 180r/min.Minimal medium composition: Na
2HPO
412H
2O, 7g/L; KH
2PO
4, 1g/L; CaCl
22H
2O, 10mg/L; FeCl
3, 2mg/L; MgSO
47H
2O, 20mg/L.When bacterium liquid becomes muddy, oil of mirbane detect less than, transfer next time; Cultivate through domestication constantly, finally obtaining with oil of mirbane is the stable bacterium liquid of sole carbon, nitrogen, the energy.To stablize the dilution of bacterium liquid, on the solid medium that with oil of mirbane is sole carbon, nitrogen, the energy, be coated with.Solid culture based component: Na
2HPO
412H
2O, 7g/L; KH
2PO
4, 1g/L; CaCl
22H
2O, 10mg/L; FeCl
3, 2mg/L; MgSO
47H
2O, 20mg/L; Oil of mirbane, 200mg/L, agar 2%.This solid medium is put into illumination box, 30 ℃, cultivated 4-5 days, choose single bacterium colony; Separating a strain is the Streptomycesalbidoflhaving of sole carbon, nitrogen, the energy with oil of mirbane; And this bacterial strain is preserved in " China Committee for Culture Collection of Microorganisms's common micro-organisms " center ", its preserving number CGMCC NO.1759 on July 18th, 2006.
The appropriate pH value of this Streptomycesalbidoflhaving is 7.0, is fit to growth temperature at 30 ℃.
The 16S rDNA amplification of Streptomycesalbidoflhaving provided by the invention:
By the 16S rDNA of this bacterial strain that increases, having obtained length is the 16S rDNA D1/D2 sequence of 1433bp.The PCR primer adopts universal primer 27f (5 '-GAGTTTGATC (AC) TGGCTCAG-3 ') and the 1492r (5 '-TACGG (CT) TACCTTGTTACGACTT-3 ') of 16S rDNA.(GeneAmp, PCR System9700) carries out amplified reaction with the PCR instrument.The reaction system cumulative volume is 25 μ l: ultrapure water 16.2 μ l, and 10xBuffer 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 do to get 2 μ l in reaction system after certain dilution.94 ℃ of pre-sex change 5min, through 30 circulations, wherein, and 98 ℃ of sex change 20s, 55 ℃ of annealing 40s, 72 ℃ are extended 1.5min, extend 7min at 72 ℃ again after 30 circulations, 1% agarose gel electrophoresis, EB dyeing back ultraviolet detection.Carry out homology relatively by the Blast program, the homology that shows this bacterial strain and Streptomyces albidoflavus is up to 99%.
The cell liquid culture of preparation Streptomycesalbidoflhaving:
Picking is the Streptomycesalbidoflhaving list bacterium colony on the solid medium of sole carbon, nitrogen, the energy with oil of mirbane, what sterilization was housed is in the liquid nutrient medium of sole carbon, nitrogen, the energy with oil of mirbane, wherein the concentration of oil of mirbane is 100mg/l, in 30 ℃, pH 7.0,180r/min, carry out aerobic cultivation 60-72 hour, getting cultured bacterium liquid 1ml is seeded in the minimal medium of 100ml nitrobenzene-containing (200mg/l), 30 ℃, pH 7.0,180r/min, carry out aerobic cultivation 72 hours, obtain the cell liquid culture of Streptomycesalbidoflhaving.
The immobilized cell of preparation Streptomycesalbidoflhaving:
[1] adopts the surface adsorption immobilization technology;
[2] patented product of selecting for use Dalian Landa Biological Environmental Technology Co., Ltd. to provide, DW-22 type macroporous netlike carrier is as immobilization material, and table 3 has provided the performance of carrier;
Table 3 DW-22 type macroporous netlike carrier property
| Model | Specific surface area (m 2/m 3) | Biological charge capacity (g microorganism/L water) | Wet tap density (g/cm behind the load microorganism 3) |
| DW-22 | 1.6～2.2×10 5 | 18～40 | 0.9～1.1 |
[3] add the 100ml minimal medium in the 250ml Erlenmeyer flask, add oil of mirbane again as sole carbon, nitrogen, the energy, wherein the concentration of oil of mirbane is 200ml;
[4] DW-22 type carrier is directly added in the Erlenmeyer flask of above-mentioned [3] 121 ℃ of 20min that sterilize down; Wherein, the carrier dosage is 24% (volume ratio), and the carrier dosage is the carrier bulk of adding and the ratio of nutrient solution volume;
[5] the cell liquid culture of inoculation 1ml Streptomycesalbidoflhaving in 30 ℃, pH 7.0,160r/min, carries out aerobic cultivation;
[6] when oil of mirbane is degraded fully, adhered to a large amount of Streptomycesalbidoflhaving cells on the macroporous netlike carrier, Streptomycesalbidoflhaving is fixed on the macroporous netlike carrier;
[7] adhered to the macroporous netlike carrier of Streptomycesalbidoflhaving cell with aseptic normal saline flushing, washed altogether 3 times, this macroporous netlike carrier has been immersed in the physiological saline, 4 ℃ of storages, standby.
Embodiment 2
Among the present invention, the application of Streptomycesalbidoflhaving p-nitrophenyl aerobic degradation research, its step is as follows:
[1] add the 100ml minimal medium in the 250ml Erlenmeyer flask, the oil of mirbane that adds 400mg/L is again sterilized down for 121 ℃ as sole carbon, nitrogen, the energy;
[2] the Streptomycesalbidoflhaving cell liquid culture with embodiment 1 preparation adds in the substratum of above-mentioned [1], 30 ℃, 180r/min, and aerobic cultivation, Fig. 1 is the growth and the nitrobenzene degradation situation of Streptomycesalbidoflhaving.
Embodiment 3
Among the present invention, Streptomycesalbidoflhaving is to the application of pyridine carboxylic acid aerobic degradation research, and its step is as follows:
[1] add the 100ml minimal medium in the 250ml Erlenmeyer flask, the pyridine carboxylic acid that adds 3000mg/L is sterilized down for 121 ℃ as sole carbon, nitrogen, the energy;
[2] the Streptomycesalbidoflhaving cell liquid culture with embodiment 1 preparation adds in the substratum of above-mentioned [1], 30 ℃, 180r/min, and aerobic cultivation, Fig. 2 is the growth and the pyridine carboxylic acid degraded situation of Streptomycesalbidoflhaving.
Embodiment 4
Among the present invention, the application of Streptomycesalbidoflhaving p-nitrophenyl aerobic degradation research under high salinity, its step is as follows:
[1] preparing the sodium-chlor mass concentration respectively is 2%, 3%, 5% and 7% minimal medium, and the oil of mirbane that adds 400mg/L is as sole carbon, nitrogen, the energy, 121 ℃ of sterilizations down;
[2] the Streptomycesalbidoflhaving cell liquid culture with embodiment 1 preparation adds in the substratum of above-mentioned [1], 30 ℃, 180r/min, and aerobic cultivation, Fig. 3 is growth and the nitrobenzene degradation situation of Streptomycesalbidoflhaving under different salinity.
Embodiment 5
Among the present invention, Streptomycesalbidoflhaving when the coexistence of aniline and oil of mirbane, the application of p-nitrophenyl aerobic degradation research, its step is as follows:
[1] at the minimal medium of 100ml, add the oil of mirbane of 400mg/L, add the aniline of different concns more respectively, aniline concentration changes between 25-100mg/L, 121 ℃ of sterilizations down;
[2] the Streptomycesalbidoflhaving cell liquid culture with embodiment 1 preparation adds in the substratum of above-mentioned [1], and 30 ℃, 180r/min, aerobic cultivation 144h, Fig. 4 are growth and the nitrobenzene degradation situation of Streptomycesalbidoflhaving when aniline and oil of mirbane coexistence.
Embodiment 6
Among the present invention, Streptomycesalbidoflhaving when the coexistence of phenol and oil of mirbane, the application of p-nitrophenyl aerobic degradation research, its step is as follows:
[1] at the minimal medium of 100ml, add the oil of mirbane of 400mg/L, add the phenol of different concns more respectively, phenol concentration changes between 50-250mg/L, 121 ℃ of sterilizations down;
[2] the Streptomycesalbidoflhaving cell liquid culture with embodiment 1 preparation adds in the substratum of above-mentioned [1], and 30 ℃, 180r/min, aerobic cultivation 144h, Fig. 5 are growth and the nitrobenzene degradation situation of Streptomycesalbidoflhaving when phenol and oil of mirbane coexistence.
Embodiment 7
Among the present invention, the application of Streptomycesalbidoflhaving immobilized cell p-nitrophenyl aerobic degradation research, its step is as follows:
[1] liquid nutrient medium of configuration embodiment 4 described different salinity;
[2] the configuration embodiment 5 described liquid nutrient mediums that contain different concns aniline;
[3] the configuration embodiment 6 described liquid nutrient mediums that contain different concns phenol;
[4] embodiment 1 described Streptomycesalbidoflhaving immobilized cell is added in the substratum of above-mentioned [1], [2], [3] 30 ℃, 160r/min, aerobic cultivation respectively; Table 4 has been listed the degraded of Streptomycesalbidoflhaving immobilized cell p-nitrophenyl under different salinity, and table 5 has been listed the degraded of Streptomycesalbidoflhaving immobilized cell p-nitrophenyl when phenol or aniline and oil of mirbane coexistence; Be that the degradation rate of Streptomycesalbidoflhaving immobilized cell 216h p-nitrophenyl is 69.43% under 5% the condition in salinity, and Streptomycesalbidoflhaving cell liquid culture only is 33.16% at the degradation rate of 216h p-nitrophenyl; When phenol concentration was 200mg/L, the degradation rate of Streptomycesalbidoflhaving immobilized cell 144h p-nitrophenyl was 92.67%, and Streptomycesalbidoflhaving cell liquid culture only is 45.23% at the degradation rate of 144h p-nitrophenyl; When aniline concentration was 50mg/L, the degradation rate of Streptomycesalbidoflhaving immobilized cell 144h p-nitrophenyl was 63.28%%, and Streptomycesalbidoflhaving cell liquid culture is 12.31% at the degradation rate of 144h p-nitrophenyl; The result shows with the cell liquid culture and compares that the salt tolerance and the resistance to poison of immobilized cell all increase.
The degraded of table 4 Streptomycesalbidoflhaving immobilized cell p-nitrophenyl under different salinity
| Salinity (%) | Water inlet (mg/L) | Oil of mirbane water outlet (mg/L) | Clearance (%) | Degradation time (h) |
| 2 3 5 7 | 414.49 401.13 407.23 400.56 | 7.30 43.64 124.49 206.47 | 98.24 89.12 69.43 23.49 | 144 216 216 216 |
The degraded of table 5 Streptomycesalbidoflhaving immobilized cell p-nitrophenyl when phenol or aniline exist
| Phenol (mg/L) | Oil of mirbane | Aniline (mg/L) | Oil of mirbane | ||||
| Water inlet (mg/L) | Water outlet (mg/L) | Clearance (%) | Water inlet (mg/L) | Water outlet (mg/L) | Clearance (%) | ||
| 50 100 150 200 | 399.53 411.26 401.89 400.21 | 5.23 11.23 8.85 29.34 | 98.69 97.27 97.82 92.67 | 25 50 75 100 | 405.56 407.09 396.53 404.15 | 4.83 149.48 268.97 343.24 | 98.81 63.28 32.17 15.07 |
Embodiment 8
Among the present invention, the Streptomycesalbidoflhaving immobilized cell is to the application of pyridine carboxylic acid aerobic degradation research, and its step is as follows:
[1] add the 100ml minimal medium in the 250ml Erlenmeyer flask, the pyridine carboxylic acid that adds 3000mg/L is sterilized down for 121 ℃ as sole carbon, nitrogen, the energy;
[2] the Streptomycesalbidoflhaving immobilized cell with embodiment 1 preparation adds in the substratum of above-mentioned [1], 30 ℃, 160r/min, and aerobic cultivation, degrade fully time of pyridine carboxylic acid of immobilized cell is 47h.
＜110〉Dalian University of Technology's environment and life institute
＜120〉have the Streptomycesalbidoflhaving of new metabolic characteristics and the application in biological degradation thereof
＜160〉the sequence sum 1
<211>1433bp
<212>DNA
＜213〉Streptomycesalbidoflhaving (Streptomyces albidoflavus)
<220>
<221>gene
＜120〉Streptomycesalbidoflhaving 16S rDNA sequence
1 gtgaatgcgg cgtgcttacc atgcaagtcg aacgatgaac cgctttcggg cggggattag
61 tggcgaacgg gtgagtaaca cgtgggcaat ctgccctgca ctctgggaca agccctggaa
121 acggggtcta ataccggata tgactgtcca tcgcatggtg gatggtgtaa agctccggcg
181 gtgcaggatg agcccgcggc ctatcagctt gttggtgagg tagtggctca ccaaggcgac
241 gacgggtagc cggcctgaga gggcgaccgg ccacactggg actgagacac ggcccagact
301 cctacgggag gcagcagtgg ggaatattgc acaatgggcg aaagcctgat gcagcgacgc
361 cgcgtgaggg atgacggcct tcgggttgta aacctctttc agcagggaag aagcgaaagt
421 gacggtacct gcagaagaag cgccggctaa ctacgtgcca gcagccgcgg taatacgtag
481 ggcgcaagcg ttgtccggaa ttattgggcg taaagagctc gtaggcggct tgtcacgtcg
541 gttgtgaaag cccggggctt aaccccgggt ctgcagtcga tacgggcagg ctagagttcg
601 gtaggggaga tcggaattcc tggtgtagcg gtgaaatgcg cagatatcag gaggaacacc
661 ggtggcgaag gcggatctct gggccgatac tgacgctgag gagcgaaagc gtggggagcg
721 aacaggatta gataccctgg tagtccacgc cgtaaacggt cggcactagg tgtgggcaac
781 attccacgtt gtccgtgccg cagctaacgc attaagtgcc ccgcctgggg agtacggccg
841 caaggctaaa actcaaagga attgacgggg gcccgcacaa gcggcggagc atgtggctta
901 attcgacgca acgcgaagaa ccttaccaag gcttgacata caccggaaac gtctggagac
961 aggcgccccc ttgtggtcgg tgtacaggtg gtgcatggct gtcgtcagct cgtgtcgtga
1021 gatgttgggt taagtcccgc aacgagcgca acccttgtcc cgtgttgcca gcaggccctt
1081 gtggtgctgg ggactcacgg gagaccgccg gggtcaactc ggaggaaggt ggggacgacg
1141 tcaagtcatc atgcccctta tgtcttgggc tgcacacgtg ctacaatggc cggtacaatg
1201 agctgcgata ccgcgaggtg gagcgaatct caaaaagccg gtctcagttc ggattggggt
1261 ctgcaactcg accccatgaa gtcggagtcg ctagtaatcg cagatcagca ttgctgcggt
1321 gaatacgttc ccgggccttg tacacaccgc ccgtcacgtc acgaaagtcg gtaacacccg
1381 aagccggtgg cccaacccct tgtgggaggg agcttcgaag gtgactgcat cca
Claims (7)
1, has the Streptomycesalbidoflhaving of new metabolic characteristics and the application in biological degradation thereof, it is characterized in that, Streptomycesalbidoflhaving is preserved in " China Committee for Culture Collection of Microorganisms's common micro-organisms " center "; preserving number CGMCC No.1759, classification called after Streptomyces albidoflavus on July 18th, 2006.
2, according to claim 1 have the Streptomycesalbidoflhaving of new metabolic characteristics and an application in biological degradation thereof, it is characterized in that, from Dalian Dye Factory aeration tank, get the 50ml active sludge, the trisodium phosphate of adding 0.2% is as separating flocculation agent, after vibration 5-10min smashes the active sludge flco on the vibrator, get the 5ml active sludge, adding contains shaking in the bottle of 95ml minimal medium, add oil of mirbane again as sole carbon, nitrogen, the energy, wherein the concentration of oil of mirbane is 200mg/L, carries out shaking table and cultivates, 30 ℃ of temperature, rotating speed 180r/min; When bacterium liquid becomes muddy, transfer, obtaining with oil of mirbane is the stable bacterium liquid of sole carbon, nitrogen, the energy; To stablize bacterium liquid dilution, on the solid medium that with oil of mirbane is sole carbon, nitrogen, the energy, be coated with, this solid medium will be put into illumination box, 30 ℃, cultivated 4-5 days, choose single bacterium colony; Separate a strain and be with oil of mirbane sole carbon, nitrogen, the energy Streptomycesalbidoflhaving.
3, claim 1 is described has the Streptomycesalbidoflhaving of new metabolic characteristics and an application in biological degradation thereof, it is characterized in that when oil of mirbane was sole carbon, nitrogen, the energy, the maximum tolerated concentration of Streptomycesalbidoflhaving p-nitrophenyl reached 400mg/L; The total clearance of TOC reaches 99.12%; Streptomycesalbidoflhaving can be degraded as the waste water of biological reinforced preparation p-nitrophenyl concentration≤400mg/L.
4, claim 1 is described has the Streptomycesalbidoflhaving of new metabolic characteristics and an application in biological degradation thereof, it is characterized in that, when oil of mirbane is that sole carbon, nitrogen, the energy and sodium chloride concentration are 0-50mg/L, Streptomycesalbidoflhaving is degrading nitrobenzene effectively; Streptomycesalbidoflhaving can be degraded to the waste water of salinity≤50mg/L, nitro phenenyl concentration≤400mg/L as biological reinforced preparation.
5, claim 1 is described has the Streptomycesalbidoflhaving of new metabolic characteristics and an application in biological degradation thereof, it is characterized in that, when oil of mirbane and aniline coexistence, aniline concentration is 0-50mg/L, and Streptomycesalbidoflhaving is degrading nitrobenzene effectively; Streptomycesalbidoflhaving can be as biological reinforced preparation to aniline and oil of mirbane coexistence, and the waste water of aniline concentration≤50mg/L, nitro phenenyl concentration≤400mg/L is degraded.
6, claim 1 is described has the Streptomycesalbidoflhaving of new metabolic characteristics and an application in biological degradation thereof, it is characterized in that, when oil of mirbane and phenol coexistence, phenol concentration is 0-200mg/L, the effective degrading nitrobenzene of Streptomycesalbidoflhaving; Streptomycesalbidoflhaving can be as biological reinforced preparation Pyrogentisinic Acid and oil of mirbane coexistence, and the waste water of phenol concentration≤200mg/L, nitro phenenyl concentration≤400mg/L is degraded.
7, claim 1 is described has the Streptomycesalbidoflhaving of new metabolic characteristics and an application in biological degradation thereof, it is characterized in that, when pyridine carboxylic acid is sole carbon, nitrogen, the energy, Streptomycesalbidoflhaving reaches 3000mg/L to the maximum tolerated concentration of pyridine carboxylic acid; The total clearance of TOC reaches 98.37%; Streptomycesalbidoflhaving can be degraded to the waste water of pyridine carboxylic acid concentration≤3000mg/L as biological reinforced preparation.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102515366A (en) * | 2011-12-23 | 2012-06-27 | 重庆文泰节能环保科技有限公司 | Microbiological degradation method for nitrobenzene industrial wastewater |
| CN103184184A (en) * | 2013-04-24 | 2013-07-03 | 牛赡光 | Streptomyces albidoflavus and applications thereof |
| CN105540997A (en) * | 2015-12-12 | 2016-05-04 | 常州大学 | Method for treating nitrobenzene wastewater |
| CN105886428A (en) * | 2016-04-05 | 2016-08-24 | 中国科学院微生物研究所 | Streptomyces albidoflavus and applications thereof in microbial fertilizers |
| CN106119159A (en) * | 2016-06-29 | 2016-11-16 | 西安交通大学 | The comamonas of one high-efficiency degradation pyridine carboxylic acid and application thereof |
| CN106906171A (en) * | 2017-05-04 | 2017-06-30 | 陕西枫丹百丽生物科技有限公司 | A kind of preparation method of apple tree canker biocontrol agent |
| CN106947721A (en) * | 2017-04-26 | 2017-07-14 | 北京农学院 | One plant is killed nematode Streptomycesalbidoflhaving and its application |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102515366A (en) * | 2011-12-23 | 2012-06-27 | 重庆文泰节能环保科技有限公司 | Microbiological degradation method for nitrobenzene industrial wastewater |
| CN102515366B (en) * | 2011-12-23 | 2013-07-10 | 重庆文泰节能环保科技有限公司 | Microbiological degradation method for nitrobenzene industrial wastewater |
| CN103184184A (en) * | 2013-04-24 | 2013-07-03 | 牛赡光 | Streptomyces albidoflavus and applications thereof |
| CN103184184B (en) * | 2013-04-24 | 2014-11-19 | 牛赡光 | Streptomyces albidoflavus and applications thereof |
| CN105540997A (en) * | 2015-12-12 | 2016-05-04 | 常州大学 | Method for treating nitrobenzene wastewater |
| CN105886428A (en) * | 2016-04-05 | 2016-08-24 | 中国科学院微生物研究所 | Streptomyces albidoflavus and applications thereof in microbial fertilizers |
| CN105886428B (en) * | 2016-04-05 | 2019-07-16 | 中国科学院微生物研究所 | One plant of Streptomycesalbidoflhaving and its application in microbial manure |
| CN106119159A (en) * | 2016-06-29 | 2016-11-16 | 西安交通大学 | The comamonas of one high-efficiency degradation pyridine carboxylic acid and application thereof |
| CN106119159B (en) * | 2016-06-29 | 2019-07-23 | 西安交通大学 | The comamonas of one high-efficiency degradation pyridine carboxylic acid and its application |
| CN106947721A (en) * | 2017-04-26 | 2017-07-14 | 北京农学院 | One plant is killed nematode Streptomycesalbidoflhaving and its application |
| CN106906171A (en) * | 2017-05-04 | 2017-06-30 | 陕西枫丹百丽生物科技有限公司 | A kind of preparation method of apple tree canker biocontrol agent |
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