CN107245463A - One plant of heterotrophic nitrification aerobic denitrifying arthrobacterium and its application - Google Patents
One plant of heterotrophic nitrification aerobic denitrifying arthrobacterium and its application Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/06—Arthrobacter
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C—CHEMISTRY; METALLURGY
- 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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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/38—Organic compounds containing nitrogen
Abstract
The invention discloses one plant of heterotrophic nitrification aerobic denitrifying arthrobacterium and its application in waste water is denitrogenated.The bacterial strain is arthrobacterium（Arthrobactersp.）WZUF01, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and collection numbering of registering on the books is CGMCC NO.14012.The bacterial strain is used to go to denitrogenate the nitrogen of sewage, has pH and Acclimation temperature scope that comparison is wide and than relatively low carbon-nitrogen mass ratio, and be resistant to the high dissolved oxygen of comparison.The bacterial strain is resistant to the ammonium of high concentration, with processing NH4 +Animal farm wastewater after anaerobic digestion of the N concentration more than 300 mg/L, result meets livestock and poultry breeding industry pollutant emission standard（GB18596‑2001）, and final product is nitrogen.The bacterial strain has good ability of aggregation.In a word, arthrobacterium WZUF01 has very big application potential in the nitrogen of actual waste water is removed.
Description
Technical field
The invention belongs to technical field of environmental microorganism, and in particular to one plant of heterotrophic nitrification-aerobic denitrification arthrobacterium with
And its application in the nitrogen for removing sewage.
Background technology
Nitrogen is become increasingly conspicuous in recent years to the pollution problem that environment is caused, and its harmfulness is also increasingly recognized and again
Depending on.As ammonia nitrogen, nitrate nitrogen and nitrite nitrogen are possible to be converted into the nitrosamine of carcinogenic, mutagenesis and teratogenesis；And for example nitrogen
Flow into water body and cause body eutrophication, cause water quality deterioration to be degenerated down to lake.Biological denitrificaion has high treating effect, processing
Process stabilization is reliable, the advantage of convenient operation and management etc. and be used widely.Traditional biological denitrificaion is by Autotrophic nitrification bacterium
The denitrification of nitrification and anaerobic denitrifying bacterium is completed.
In the 1980s, Robertson and Kuenen is isolated in sulphur removal and denitrification processing system
Thiosphaera pantotropha (are now renamed as Paracoccus denitrificans Paracoccus denitrifications), and it can
Breathed simultaneously by the use of nitrate and oxygen as terminal electron acceptor, they are referred to as aerobic denitrification.By more than 30 years both at home and abroad
The numerous studies of scholar, have affirmed the presence of aerobic denitrifying bacteria, and most of aerobic denitrifying bacterias performance Heterotrophic nitrification
Characteristic.Compared with autotrophic type nitrifier, the growth rate of nitrification bacteria is fast, cell yield it is high, it is necessary to dissolved oxygen concentration
It is low, it is resistant to sour environment and activity is high, and the nitrogen compound of various forms can be metabolized, while improving COD clearance.It is good
It is theoretical that traditional biological denitrificaion has been broken in the discovery of oxygen denitrification and heterotrophic nitrification so that simultaneous nitrification and denitrification turns into can
Can, operating cost can be not only reduced, operation cumbersome in technique is reduced, Autotrophic nitrification bacterium institute can also be expanded not treatable
Water quality scope, new approaches are provided for the biological removal of nitrogen of waste water.
Domestic and international many scholars isolate many heterotrophic nitrification-aerobic denitrification bacterium from different ecological environments, have studied
It denitrogenates characteristic and application potential.Wherein there is low emission NO, N2The heterotrophic nitrification-aerobic denitrification bacterium of O abilities is considered as
Whether there is the matter of utmost importance of practical application potentiality.Most of aerobic denitrifying bacterias, denitrifying the last two steps are NO → N2O→
N2It is sensitive, therefore larger amount of NO and N to oxygen2O can be released in aerobic nitrate/nitrate reductase stage.
Certainly, separation also obtains low emission NO, N2The report of the heterotrophic nitrification-aerobic denitrification bacterium of O abilities.Such as Maosheng
Zheng etc. from handle it is nitrogenous up to 200mg/L percolate laboratory scale BAF in be separated to 1 plant
The NO and N of aerobic denitrification stage emission low amounts2O Pseudomonas stutzeri PCN-1, can be with synthetic wastewater
Nitrate, nitrite, N2O is good denitrification substrate, and the average denitrification rates of three are respectively 11.66mg/L/
H, 12.80mg/L/h and 9.67mg/L/h；The NO and N of aerobic denitrification are carried out by substrate of nitrate2O accumulation is respectively lower than
0.003% and 0.33%；The NO and N of aerobic denitrification are carried out by substrate of nitrite2O accumulation is respectively lower than 0.006% He
0.29%；Further investigation revealed that denitrification gene nirS, cnorB and nosZ coordinate expression cause low NO and N2O is arranged
High-volume (Maosheng Zheng, et al, Bioresource Technology, 2014,162:80-88).For another example Zhuang
Shi etc. separates the heterotrophic nitrification-aerobic denitrification Paracoccus versutus LYM obtained from the sludge of seabed, can have
The nitrate nitrogen (about 400mg/L) more than 95% is transformed into gaseous state denitration product through peroxynitrite again under the conditions of oxygen, maximum is gone
Except rate reaches 33mg/L/h；When using ammonium as only nitrogen source, average removal rate and clearance be respectively 11.4mg/L/h and
75.4%, other nitrogen compounds such as azanol, nitrite, nitrate only have a small amount of accumulation, and its maximum accumulation is respectively
0.045mg/L, 0.048mg/L and 4mg/L；N in the aerobic denitrification stage2For primary product, do not detect any as temperature
The nitrogen oxides of room gas and ozone depleting substance presence (Zhuang Shi, et al, Bioresource Technology,
2013,148:144-148)。
Up to now, heterotrophic nitrification-aerobic denitrification mainly also rests on the laboratory exploratory stage, for the processing of waste water nitrogen
The report of practice is few.In view of Heterotrophic nitrification-aerobic denitrifying bacteria that different ecological environment is obtained, its physiological and ecological characteristic is each
It is different, denitrogenate characteristic and to denitrogenate ability also different, searching denitrogenates that ability is strong, low emission NO, N2O excellent Heterotrophic nitrification-aerobic
Denitrifying bacterium germ plasm resource remains as the important step that Heterotrophic nitrification-aerobic denitrification is finally applied to the processing practice of waste water nitrogen.
The content of the invention
In order to solve the above-mentioned technical problem, the present invention provides one plant of heterotrophic nitrification-aerobic denitrification bacterium, and its feature is the bacterium
Strain is to separate to obtain from the activated sludge for picking up from Wenzhou District of Zhejiang Province Dong Pian sewage treatment plants using conventional separation methods, is passed through
Simultaneously sequencing result is compared by GenBank Blast for 16SrDNA sequencings, belongs to Arthrobacter category,
Homology with Arthrobacter uratoxydans (Arthrobacter uratoxydans) is 99.28%, is compiled as Arthrobacter
sp.WZUF01。
One plant of heterotrophic nitrification-aerobic denitrification arthrobacterium that the present invention is provided, the bacterial strain is arthrobacterium (Arthrobacter
Sp.) WZUF01, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and collection is registered on the books
Numbering is CGMCC NO.14012.
The present invention also provides the application of above-mentioned arthrobacterium, and the arthrobacterium WZUF01 is used to go to denitrogenate the nitrogen of sewage：Will
Arthrobacterium WZUF01 is inoculated in nitrogen sewage with except denitrification.
The arthrobacterium WZUF01 is used for the method for going to denitrogenate the nitrogen of sewage, comprises the following steps：
1) preservation strain arthrobacterium WZUF01 is inoculated in activation medium, is cultivated, and centrifugation obtains thalline, and thalline is with sterile
OD is made after water washing600For 0.9~1.0 bacteria suspension；
2) by step 1) gained bacterial suspension inoculation cultivate to denitrogenate in nitrogen sewage；
Wherein, the nitrogen sewage is to contain NH4 +、NO3 -And NO2 -One kind or its combination sewage.
Preferably, step 1) activation medium composition is：Peptone 10g, dusty yeast 5g, NaCl 5g, H2O
1000ml, pH 7.0~7.2；
Step 1) incubation time be 10~14h, temperature be 25~30 DEG C, under 150~160r/min of rotating speed cultivate.
Preferably, step 2) be by step 1) gained bacterial suspension inoculation adds contained in carbon source, carbon source in nitrogen sewage
Carbon and nitrogen sewage in the mass ratio of contained nitrogen be 4:1~10:1.
Preferably, the carbon source is sodium citrate, sodium lactate, sodium acetate, the one of which of sodium succinate or its any group
Close.
Preferably, step 2) in go to denitrogenate the cultivation temperature of the nitrogen of sewage be 15~40 DEG C, preferably 20~35 DEG C, with
NaOH or HCl adjusts pH to be 5~9.
Preferably, step 2) in, by step 1) gained bacterial suspension inoculation in nitrogen sewage under 150~200r/min of rotating speed
Cultivate to denitrogenate.
The present invention can reach following technique effect：
1st, the Arthrobacter uratoxydans (Arthrobacter for the heterotrophic nitrification-aerobic denitrification that the present invention is provided
Uratoxydans) WZUF01 is reported first for the country.
2nd, heterotrophic nitrification-aerobic denitrification arthrobacterium (Arthrobacter sp.) WZUF01 that the present invention is provided goes to denitrogenate
Suitable carbon-nitrogen mass ratio, temperature and the pH of the nitrogen of sewage are respectively 4~10:1st, 20~35 DEG C and 5~9, there is the pH that comparison is wide
With Acclimation temperature scope and than relatively low carbon-nitrogen mass ratio, and it is resistant to the high dissolved oxygen of comparison..
3rd, heterotrophic nitrification-aerobic denitrification arthrobacterium (Arthrobacter sp.) WZUF01 that the present invention is provided is resistant to
The ammonium of high concentration, under optimum conditions, it is to NH4 +NH of-N the concentration up to 600mg/L artificial nitrogen sewage4 +- N clearances and go
Removal rates are respectively 95.78% and 24.10mg/L/h.With processing NH4 +- N concentration is cultivated after exceeding 300mg/L anaerobic digestion
Field waste water, result meets livestock and poultry breeding industry pollutant emission standard (GB18596-2001), and final product is nitrogen.
4th, heterotrophic nitrification-aerobic denitrification arthrobacterium (Arthrobacter sp.) WZUF01 nitrification ammoniums that the present invention is provided
End-product be nitrogen, do not form any nitrogen oxides greenhouse gases.
5th, heterotrophic nitrification-aerobic denitrification arthrobacterium (Arthrobacter sp.) WZUF01 that the present invention is provided has good
Good ability of aggregation, its ability of aggregation grade is that between 3~4, surface hydrophobic is 59.46%.
In a word, heterotrophic nitrification-aerobic denitrification arthrobacterium (Arthrobacter sp.) WZUF01 that the present invention is provided exists
The nitrogen of actual waste water has very big application potential in removing.
Brief description of the drawings
Fig. 1 is that Arthrobacter sp.WZUF01 grow and denitrogenated process in nitrate culture-medium.
Fig. 2 is that Arthrobacter sp.WZUF01 grow and denitrogenated process in nitrite culture medium.
Fig. 3 is that Arthrobacter sp.WZUF01 grow and denitrogenated process in ammonium culture medium.
Fig. 4 is Arthrobacter sp.WZUF01 individual morphology.
Fig. 5 is to use MEGA4.1 softwares, and ortho position connection method shows bacterial strain WZUF01 and related kind 16S rDNA sequences system
System development tree, the similarity for carrying out 1000 times is computed repeatedly, and tree node is developed in figure and only shows that Bootstrap values are more than 50% number
Value.
Fig. 6 is Arthrobacter sp.WZUF01 except animal farm wastewater TN, NH after the anaerobic digestion of addition sodium acetate4 +-N
With the process of growth.In figure (1)：Control group；(2)：Treatment group.
Fig. 7 is Arthrobacter sp.WZUF01 except animal farm wastewater NO after the anaerobic digestion of addition sodium acetate3 --N、
NO2 -- N and COD processes.In figure (1)：Control group；(2)：Treatment group.
Bacterial strain preservation
The Arthrobacter sp.WZUF01 of the present invention, have been preserved in China Committee for Culture Collection of Microorganisms general
Logical microorganism center, preservation address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, collection numbering of registering on the books is
CGMCC NO.14012, preservation from date is on April 11st, 2017.
Embodiment
The invention will be further described with specific embodiment below in conjunction with the accompanying drawings, so that those skilled in the art can be with
More fully understand the present invention and can be practiced, but illustrated embodiment is not as a limitation of the invention.
The present invention provides one plant of heterotrophic nitrification-aerobic denitrification bacterium, and its feature is that the bacterial strain is using enrichment culture, just
The processes such as sieve, secondary screening are separated from the activated sludge for picking up from the eastern piece sewage disposal plant aeration tank of Wenzhou District of Zhejiang Province and obtained, through 16SrDNA
Simultaneously sequencing result is compared by GenBank Blast for sequencing, belongs to Arthrobacter sp., with uric acid
The homology for aoxidizing arthrobacterium (Arthrobacter uratoxydans) is 99.28%, is compiled as Arthrobacter
sp.WZUF01
Embodiment one：Heterotrophic nitrification-aerobic denitrification arthrobacterium WZUF01 separation
(1) primary dcreening operation
The activated sludge 5ml for picking up from the eastern piece sewage disposal plant aeration tank in Wenzhou is taken to be inoculated in equipped with 100ml enriched mediums
(sodium succinate 5g, NaNO3 1g、KH2PO 4 1g、K2HPO4 1g、(NH4)2SO4 0.25g、MgSO4·7H2O 0.2g, yeast
Powder 1g, H2In O 1000ml, the 250ml conical flasks of pH 7.0~7.2), shaken cultivation 3 days under 25 DEG C, 150rpm；Take 10ml
Enrichment culture liquid is transferred in fresh enriched medium and cultivated under the same conditions, is so repeated 5 times；Enrichment culture liquid is taken to pass through
Coating is inoculated in bromthymol blue (Bromothymol Blue) flat board (sodium succinate 5g, NaNO after appropriate dilution3 1g、KH2PO4 1g、K2HPO4 1g、MgSO4·7H2O 1g、CaCl2·2H2O 0.2g、FeSO4·7H2O 0.05g, asparagine 1g, bromine hundred
In phenol indigo plant 1ml (with 1% ethanol prepare), agar 18g, H2On O 1000ml, pH 7.0~7.2), cultivated 2-3 days at 25 DEG C,
Selecting the single bacterium colony line switching bromthymol blue (Bromothymol Blue) with the blue ring of light, flat board is under the same conditions
Culture, until being the preservation at 4 DEG C after switching LB inclined-plane cultures after pure culture through microscopy.Primary dcreening operation obtains 20 plants of pure cultures altogether
Thing.
(2) secondary screening
The slant preservation bacterium of 20 plants of pure cultures of primary dcreening operation acquisition is taken to be inoculated in respectively equipped with 100ml LB culture medium (albumen
Peptone 10g, dusty yeast 5g, NaCL 5g, H2O 1000ml, pH 7.0-7.2) 250ml conical flasks in, under 25 DEG C, 150rpm
12h is cultivated, thalline is collected after centrifuging 15min under 5000rpm, OD is configured to after washing 3 times with sterile distilled water600=0.9~
1.0 bacteria suspension, the bacteria suspension of every plant of pure culture takes 10ml to be inoculated in respectively equipped with 200ml nitrate culture-mediums, Asia
In the 500ml conical flasks of nitrate culture-medium and ammonium culture medium, the shaken cultivation under 25 DEG C, 160r/min.Culture medium it is basic
Constitute and be：Sodium succinate 5g, KH2PO 4 1g、K2HPO4 1g、MgSO4·7H2O 1g、CaCl2·2H2O 0.2g、FeSO4·
7H2O 0.05g、H2O 1000ml, pH 7.0~7.2, nitrate culture-medium add NaNO30.91g/L is (using nitrate as matrix
Denitrification), nitrite culture medium add NaNO20.73g/L (using nitrite as the denitrification of matrix),
Ammonium culture medium adds NH4CL 0.57g/L (using ammonium chloride as the nitrification of matrix), nitrate culture-medium, nitrite culture
The mass ratio that nitrogen is 150mg/L, C and N in base and ammonium culture medium is 10:1.Timing sampling determines biomass and nitric acid
Salt, nitrite, ammonium, azanol, the change in concentration of albumen and succinic acid.
The pure culture obtained according to 20 plants of primary dcreening operations aoxidizes NO in nitrate culture-medium, nitrite culture medium respectively3 -、
NO2 -Its aerobic denitrification is determined with intermediate product type and change in concentration, NH is aoxidized in ammonium culture medium4 +, intermediate product
Type and change in concentration and the change in concentration of butanedioic acid determine its heterotrophic nitrification, as a result screen 5 plants of NO3 -、NO2 -、NH4 +
Clearance is all higher than 80% heterotrophic nitrification-aerobic denitrification bacterium.Wherein biomass estimation OD600, NO3 --N、NO2 -- N and NH4 +-
Disulfonic acid phenol AAS, N- (1- naphthyls)-ethylenediamine photometry and reagent colorimetric method is respectively adopted in N measure,
Protein determination uses Folin- phenol methods, azanol determine using itself and excessive 8-hydroxyquinoline formed stable 5,8- quinoline quinones-
5- (8- hydroxyl -5- quinoline acid imide) quantitative AAS (Frear DS, Anal.Chem., 1955,27:1664-
1665), succinate is determined using KSUCC amber acidity tests kits (Michal G H, Anal.Chem., 1976,279:
137-138),
Screen 5 plants of heterotrophic nitrification-aerobic denitrification bacterium are subjected to nitrogen balance tests, method be by 5 plants of heterotrophic nitrifications-
Aerobic denitrifying bacteria is inoculated in equipped with 100ml LB culture mediums (peptone 10g, dusty yeast 5g, NaCL 5g, H respectively2O
1000ml, pH 7.0-7.2) 250ml conical flasks in, cultivate 12h under 25 DEG C, 150r/min, under 5000rpm centrifuge
Thalline is collected after 15min, OD is configured to after washing 3 times with sterile distilled water600=0.9~1.0 bacteria suspension, every plant of heterotrophism nitre
The bacteria suspension of change-aerobic denitrifying bacteria respectively takes 10ml to be inoculated in respectively equipped with 200ml nitrate culture-mediums, nitrite culture
In the 500ml conical flasks of base and ammonium culture medium, then with purity oxygen lead to 10min after with rubber septum seal conical flask, 25 DEG C,
Sealing culture 18h under 150r/min.Culture medium basic composition is：Sodium succinate 5g, KH2PO 4 1g、K2HPO4 1g、
MgSO4·7H2O 1g、CaCl2·2H2O 0.2g、FeSO4·7H2O 0.05g、H2O 1000ml, pH 7.0~7.2, nitrate
Culture medium adds NaNO30.91g/L (using nitrate as the denitrification of matrix), nitrite culture medium adds NaNO2
0.73g/L (using nitrite as the denitrification of matrix), ammonium culture medium adds NH4CL 0.57g/L are (using ammonium chloride as base
The nitrification of matter), nitrogen is 150mg/L, C and N quality in nitrate culture-medium, nitrite culture medium and ammonium culture medium
Than being 10:1.The change of culture medium nitrogen, thalline nitrogen and headroom nitrogen and oxygen before and after analysis culture, to determine what is removed
NO3 -、NO2 -、NH4 +Whether the nitrogen compound of headroom is changed into, and screening final product is anti-for the heterotrophic nitrification-aerobic of nitrogen
Nitrifier.As a result 1 plant of NO is obtained3 -、NO2 -、NH4 +It is anti-that clearance is all higher than the heterotrophic nitrification-aerobic that 90%, end-product is nitrogen
Nitrifier, compiles as WZUF01.The oxygen and nitrogen content wherein sealed in the top gas of culture uses GC-14B gas-chromatographies
Instrument is determined, and thalline nitrogen is determined using micro-Kjeldahl.
Fig. 1, Fig. 2 and Fig. 3 be respectively during secondary screening bacterial strain WZUF01 nitrate culture-medium, nitrite culture medium and
The experimental result that growth and nitrogen concentration change in ammonium culture medium, Tables 1 and 2 is bacterial strain WZUF01 nitrogen balance test results.
The closed culture 18h of the bacterial strain WZUF01 of table 1 nitrogen balance test result (mg/L)
The closed culture 18h headrooms nitrogen of the bacterial strain WZUF01 of table 2 and oxygen change (%)
It can be seen that with reference to Fig. 1, bacterial strain WZUF01 cultivates NO after 18h in nitrate culture-medium3 -- N clearances reach
97.57%, NO3 -- N removal rates are 8.21mg/L/h, and occur mainly in logarithmic phase, are synchronous with growth；Nitrite
Accumulated since 6h, 12h reaches maximum (21.89 ± 2.15mg/L), and zero is reduced to 24h；Whole process does not detect ammonium, hydroxyl
Amine and albumen；With reference to the nitrogen balance analysis shows of table 1, culture 18h wild Oryza species and thalline total nitrogen lose 54.17%, with reference to table 2
Closed culture 18h headrooms nitrogen and oxygen change, show nitrate by bacterial strain WZUF01 aerobic denitrifications be nitrous
Hydrochlorate is until nitrogen.
Understand that situations of the bacterial strain WZUF01 in nitrite culture medium is similar in nitrate culture-medium with reference to Fig. 2,
Nitrite remove occur mainly in logarithmic phase, with growth be it is synchronous, whole process do not detect ammonium, azanol, albumen and
Nitrate；Cultivate NO after 18h2 -- N is down to 4.35 ± 0.55mg/L, NO2 -- N removal rates and clearance are respectively 8.04mg/L/
H and 97.08%, understands with reference to the nitrogen balance analysis of Tables 1 and 2, wherein 52.11% nitrogen is converted into the nitrogen of headroom.
Find out with reference to Fig. 3, bacterial strain WZUF01 amine-oxides formation azanol, nitrite and nitrate in ammonium culture medium, together
When gradually reduced with the growth succinic acid concentration of bacterium, show bacterial strain WZUF01 heterotrophism；NH after 18h4 +- N is down to
2.97mg/L, NH4 +- N clearance and removal rate is respectively 98.03% and 8.22mg/L.Nitration product azanol, nitrite
Start to occur in 3h, 6h and 6h respectively with nitrate, be reduced to zero to 24h, show occur heterotrophic nitrification-aerobic denitrification；Knot
The nitrogen balance analysis of Tables 1 and 2 is closed, shows ammonium in addition to for thalline increase, remaining is 46.46% anti-by heterotrophic nitrification-aerobic
Nitrify the nitrogen for headroom.
It is molten to change by changing shaking speed in order to further prove that bacterial strain WZUF01 is to carry out aerobic denitrification
The method of oxygen detects dissolved oxygen to the denitrifying influences of bacterial strain WZUF01, when shaking speed is respectively 0,50,100 and 150 and 200r/
During min, its corresponding dissolved oxygen level is 2.35,3.55,4.94,6.18 and 7.35mg/L, dissolved oxygen level Orion-3-star
310D-01 oxygen electrodes are measured.Bacterial strain WZUF01 in aforementioned condition identical nitrite culture medium when with cultivating, NO2 -- N is gone
Except rate is respectively 10.62%, 21.89%, 64.2%, 98.74% and 97.45%, show bacterial strain WZUF denitrifications in aerobic bar
Carried out under part and higher dissolved oxygen level can be tolerated.
In order to realize that the heterotrophic nitrification-aerobic denitrification bacterium of acquisition has practical application potentiality, bacterial strain WZUF01 is determined
Ability of aggregation and surface hydrophobic.After measured, between bacterial strain WZUF01 ability of aggregation grade is 3-4, surface hydrophobic is
59.46%.
The assay method of ability of aggregation：25 DEG C in nitrate culture-medium, nitrite culture medium or ammonium culture medium,
24h nutrient solution is cultivated under 150r/min, distilled water washing thalline is used 3 times after being centrifuged through 5000r/min 15min, uses distilled water
It is made into OD600For 1.5 bacteria suspension, it is then transferred into quartz ampoule mesoscale eddies 0.5min, gently rolls 1min, then stands range estimation
Ability of aggregation；Ability of aggregation grade basis for estimation is：0, suspension is not assembled；1, there is small uniform aggregation in turbid solution；
2, there is readily visible aggregation in turbid solution；3, there is clearly supernatant in high-visible aggregation；4, almost moment appearance
Big cotton-shaped aggregation, leaves clearly supernatant (Richard A H, Appl.Environ.Microbiol., 2002,68:
3644-3650)。
The assay method of surface hydrophobic：25 DEG C in nitrate culture-medium, nitrite culture medium or ammonium culture medium,
24h nutrient solution is cultivated under 150r/min, distilled water washing thalline is used 3 times after being centrifuged through 5000r/min 15min, with pH 7.4
Phosphate buffer be made into OD6001.0 bacteria suspension, takes 1.2ml bacteria suspensions and 0.8ml bacteria suspensions (plus 0.4ml 16 respectively
Alkane) it is transferred to the OD that low aqueous phase is determined after teat glass the mesoscale eddies 2min, two-phase laminated flow 15min crossed with 5ml acid elutions600, table
Face hydrophobicity handled according to hexadecane after bacterial suspension to indigenous bacteria suspension OD600Percentage reduce calculating (Rosenberg,
FEMS Microbiol,1980,9:29-33)。
Embodiment two：Heterotrophic nitrification-aerobic denitrification arthrobacterium WZUF01 identification
Bacterial strain WZUF01 young ages cell is irregular shaft-like, normal V-shaped arrangement, shaft-like in growth course to be fractured into bead
Shape, has obvious bar, ball mechanical periodicity, G+(see Fig. 4)；It is aerobic, contact enzyme positive.
Using bacterial genomes DNA as template amplification 16SrDNA, amplification is using a pair of universal primers：Sense primer (P1)：
5 '-AGAGTTTGATCCTGGTCAGAACGAACGCT-3 ', anti-sense primer (P6)：5’-TACGGCTACCTTGTTACGACTTCAC
CCC-3 ', PCR, the purifying of PCR primer and sequencing are completed by Chinese industrial Microbiological Culture Collection administrative center, and sequencing result leads to
GenBank Blast are crossed to be compared.
The 16SrDNA of WZUF01 bacterial strains is by 1594bp base compositions, as shown in SEQ ID NO.1；Pass through GenBank
Blast is compared, and has with the 16SrDNA sequences of the Arthrobacter (Arthrobacter) in GenBank very high same
Source property, the homology with Arthrobacter uratoxydans (Arthrobacter uratoxydans) is 99.28%.Using MEGA4.1
Software, ortho position connection method shows heterotrophic nitrification-aerobic denitrification arthrobacterium WZUF01 and related kind 16S rDNA sequential systems
Development tree sees Fig. 5.
Arthrobacterium (Arthrobacter sp.) WZUF01, has been preserved in China Committee for Culture Collection of Microorganisms general
Logical microorganism center, its collection numbering of registering on the books is CGMCC NO.14012, and preservation from date is April 11 in 2017
Day.
Embodiment three：Heterotrophic nitrification-aerobic denitrification arthrobacterium WZUF01 denitrogenates characteristic
The heterotrophic nitrification-aerobic denitrification arthrobacterium WZUF01 (1.5mL cryovials melt bacterium solution) of preservation, which is inoculated in, to be equipped with
100ml LB culture mediums (peptone 10g, dusty yeast 5g, NaCL 5g, H2O 1000ml) 250ml conical flasks in, 25 DEG C,
12h is cultivated under 150r/min, thalline is collected after centrifuging 15min under 5000r/min, is washed and is prepared after 3 times with sterile distilled water
Into OD600=0.9~1.0 bacteria suspensions.
Take 5ml to be inoculated in the 250ml conical flasks equipped with 100ml culture mediums, certain temperature and rotating speed (basal temperature and
Rotating speed is respectively 25 DEG C, 150rpm) under shaken cultivation 24h, then under 5000r/min centrifuge 10min obtain supernatant, to supernatant
Liquid carries out related assays analysis.
The composition of minimal medium is：Sodium succinate 5g, KH2PO 4 1g、K2HPO4 1g、MgSO4·7H2O 1g、
CaCl2·2H2O 0.2g、FeSO4·7H2O 0.05g、H2O 1000ml, pH 7.0~7.2, are separately added into NaNO3 0.91g/L
(using nitrate as the aerobic denitrification of matrix) and NH4CL 0.57g/L (using ammonium chloride as the heterotrophic nitrification of matrix-
Aerobic denitrification), it is 10 to make the mass ratio that nitrogen is 150mg/L, C and N in culture medium:1.
NH4 +- N (or NO3 -- N) clearance/%=(supernatant NH before culture4 +- N (or NO3 -- N) concentration (mg/L)-training
Supernatant NH after supporting4 +- N (or NO3 -- N) the preceding supernatant NH of concentration (mg/L)/culture4 +- N (or NO3 -- N) concentration (mg/L).
The influence that main research carbon source, C/N, temperature, pH, rotating speed are denitrogenated to arthrobacterium WZUF01.
1st, the selection of carbon source
The sodium succinate in minimal medium is replaced with the different carbon source in table 3 respectively, the mass ratio for making C and N is 10:
1, its experimental result is as shown in table 3.Found out by table 3, optimum carbon source be sodium acetate, sodium succinate and sodium citrate, secondly
For sodium lactate.
The carbon source choice experiment result of table 3
2nd, the selection of carbon-nitrogen mass ratio (C/N)
Sodium succinate in minimal medium is replaced with sodium acetate, when experimental result is as shown in table 4 for the quality of carbon nitrogen.By
Table 4 understands that suitable carbon-nitrogen mass ratio is 4~10:1, show that heterotrophic nitrification-aerobic denitrification arthrobacterium WZUF01 need not be compared with
High carbon-nitrogen mass ratio.
The carbon-nitrogen mass ratio choice experiment result of table 4
3rd, the selection of cultivation temperature
The sodium succinate of minimal medium is replaced with sodium acetate, concentration is 2.56g/L, the mass ratio for making C and N is 5:1, temperature
The selection of degree and experimental result such as table 5.Found out by table 5, optimum temperature be 20-35 DEG C, but 15 DEG C at still have more than 60%
Clearance, show that heterotrophic nitrification-aerobic denitrification arthrobacterium WZUF01 has very wide Acclimation temperature scope.
The experimental result of the cultivation temperature of table 5 selection
4th, pH selection
The sodium succinate of minimal medium is replaced with sodium acetate, concentration is 2.56g/L, the mass ratio for making C and N is 5:1, temperature
25 DEG C of degree, the pH selections of culture medium and experimental result such as table 6.As shown in Table 6, suitable pH scopes are 5-9, show heterotrophism nitre
Change-aerobic denitrification arthrobacterium WZUF01 has very wide pH accommodations.
The experimental result of the pH of table 6 selections
5th, the selection of rotating speed is cultivated
The sodium succinate of minimal medium is replaced with sodium acetate, concentration is 2.56g/L, the mass ratio for making C and N is 5:1, temperature
25 DEG C of degree, the pH of culture medium is 7, shaking speed selection and experimental result such as table 7.Found out by table 7, suitable rotating speed is 150-
200r/min, shows that heterotrophic nitrification-aerobic denitrification arthrobacterium WZUF01 is resistant to the high dissolved oxygen level of comparison.
Table 7 cultivates the experimental result of selection of speed
Example IV：Heterotrophic nitrification-aerobic denitrification arthrobacterium WZUF01 is except ammonia nitrogen experiment under optimum condition
The heterotrophic nitrification-aerobic denitrification arthrobacterium WZUF01 (1.5mL cryovials melt bacterium solution) of preservation, which is inoculated in, to be equipped with
100ml LB culture mediums (peptone 10g, dusty yeast 5g, NaCL 5g, H2O 1000ml) 250ml conical flasks in, 25 DEG C,
12h is cultivated under 150r/min, thalline is collected after centrifuging 15min under 5000r/min, is washed and is prepared after 3 times with sterile distilled water
Into OD600=0.9~1.0 bacteria suspensions.
Take 10ml bacterial suspension inoculations in the 500ml conical flasks equipped with 200ml culture mediums, shaken under 25 DEG C, 150r/min
Culture is swung, timing sampling determines biomass, then centrifuged under 5000r/min and related assays point are carried out to supernatant after 10min
Analysis, as a result as shown in table 8.
The composition of culture medium：Sodium acetate, NH4CL、KH2PO 4 1g、K2HPO4 1g、MgSO4·7H2O 1g、CaCl2·
2H2O 0.2g、FeSO4·7H2O 0.05g、H2NH in O 1000ml, pH 7.0~7.2, culture medium4 +- N concentration is respectively 150,
300th, 600,800,1000mg/L, C and N mass ratio are 5:1.
Calculated and learnt by table 8, work as NH4 +When-N concentration is respectively 150,300,600,800 and 1000mg/L, heterotrophic nitrification-
Aerobic denitrification arthrobacterium WZUF01 is to NH4 +- N clearance is respectively 96.20%, 97.25%, 95.78%, 90.58% and
73.09%, removal rate is respectively 8.07,12.10,24.10,17.29 and 13.66mg/L/h, i.e., with NH4 +The increase of-N concentration,
Heterotrophic nitrification-aerobic denitrification arthrobacterium WZUF01 is improved therewith to its removal rate, but more than 600mg/L after, its remove speed
Rate is begun to decline again, shows the NH of excessive concentrations4 +- N shows depression effect to it；This and heterotrophic nitrification-aerobic denitrification section
Bacillus WZUF01 is in different NH4 +Biomass variety under-N concentration is consistent, and NH4 +- N removal is synchronous with growth.
It will also realize that by table 8, work as NH4 +When-N concentration is less than 600mg/L, NO2 -- N almost no accumulation (concentration ＜ 0.1mg/L),
NH4 +- N concentration is more than NO after 600mg/L2 -- N starts gradually to accumulate；And NO3 -- N concentration no matter NH4 +- N concentration is much, or
Or do not accumulate Nong Du≤1mg/L.
Embodiment five：Heterotrophic nitrification-aerobic denitrification arthrobacterium WZUF01 removes animal farm wastewater nitrogen after anaerobic digestion and tested
The heterotrophic nitrification-aerobic denitrification arthrobacterium WZUF01 (1.5mL cryovials melt bacterium solution) of preservation, which is inoculated in, to be equipped with
100ml LB culture mediums (peptone 10g, dusty yeast 5g, NaCL 5g, H2O 1000ml) 250ml conical flasks in, 25 DEG C,
12h is cultivated under 150r/min, thalline is collected after centrifuging 15min under 5000r/min, is washed and is prepared after 3 times with sterile distilled water
Into OD600=0.9-1.0 bacteria suspensions.Bacteria suspension gives up by the 5% volume ratio plant that transfers respectively after equipped with 200mL anaerobic digestions
After the 200mL anaerobic digestions of water and addition sodium acetate in the 500mL conical flasks of animal farm wastewater, under 25 DEG C, 150r/min
Culture, and cultivated under the same conditions as control with not accessing the raw wastewater of bacteria suspension respectively, timing sampling determines biomass, and
10min is centrifuged under 8000r/min and obtains supernatant progress coherent detection calculating, untill tending towards stability.Nitrogen is carried out simultaneously to put down
Weighing apparatus experiment, method is that bacteria suspension is transferred animal farm wastewater and addition after equipped with 200mL anaerobic digestions respectively by 5% volume ratio
After the 200mL anaerobic digestions of sodium acetate in the 500mL conical flasks of animal farm wastewater, rubber is used after then leading to 10min with purity oxygen
Partition seals conical flask, culture is sealed under 25 DEG C, 150r/min, culture medium nitrogen, thalline nitrogen and top are empty before and after analysis culture
Between nitrogen and oxygen change.
Raw wastewater is constituted：TN (total nitrogen) 402.89 ± 4.69mg/L, NH4 +-N 329.34±5.58mg/L、NO3 --N
35.19±1.54mg/L、NO2 --N 34.58±1.15mg/L、COD 378.42±5.87mg/L。
TN is determined clears up ultraviolet spectrophotometry (GB-11894-89) using alkaline chitinase, and COD is determined using weight chromium
Sour potassium method.
Table 9 is the effect that heterotrophic nitrification-aerobic denitrification arthrobacterium WZUF01 removes animal farm wastewater nitrogen after anaerobic digestion, table
10 remove addition 6.89g/L sodium acetates for heterotrophic nitrification-aerobic denitrification arthrobacterium WZUF01 makes sodium acetate carbon containing and total nitrogen
Mass ratio is 5:The effect of animal farm wastewater nitrogen after 1 anaerobic digestion, Fig. 6 and Fig. 7 are arthrobacterium WZUF01 except addition sodium acetate
Anaerobic digestion after animal farm wastewater nitrogen process.The arthrobacterium WZUF01 of table 9 removes the result of animal farm wastewater nitrogen after anaerobic digestion
The arthrobacterium WZUF01 of table 10 is except the result of animal farm wastewater nitrogen after the anaerobic digestion of addition sodium acetate
Contrast table 9 and table 10 are as can be seen that arthrobacterium WZUF01 is to animal farm wastewater after the anaerobic digestion of addition sodium acetate
TN, NO3 --N、NO2 --N、NH4 +- N removal effect, which is significantly higher than, to be not added with after the former anaerobic digestion of sodium acetate plant and gives up
Water.Found out by the nitrogen balance trial result of table 11, do not added after the former anaerobic digestion of sodium acetate before and after animal farm wastewater culture
Nitrogen loss 12.97%, and add nitrogen loss 63.09% before and after anaerobic digestion-rear animal farm wastewater culture of sodium acetate.Show not
Add animal farm wastewater after the former anaerobic digestion of sodium acetate and be mainly used in arthrobacterium WZUF01 growth, and add big after sodium acetate
Promote heterotrophic nitrification-aerobic denitrification greatly.With reference to table 12 closed culture after headroom nitrogen and oxygen change, show to detest
The nitrogen of animal farm wastewater is nitrogen by arthrobacterium WZUF01 heterotrophic nitrification-aerobic denitrifications after oxygen digestion.
The arthrobacterium WZUF01 of table 11 removes the nitrogen balance trial result of animal farm wastewater nitrogen after anaerobic digestion
The arthrobacterium WZUF01 of table 12 headroom nitrogen and oxygen after closed culture in animal farm wastewater after anaerobic digestion
Change (%)
It can be seen from Fig. 6 that access arthrobacterium WZUF01 cultivates 72h in animal farm wastewater after the anaerobic digestion of addition sodium acetate
Afterwards, TN and NH4 +- N is removed and tended towards stability, and is synchronous, and control group TN and NH with thalli growth4 +- N does not almost decline, NH4 +- N clearance and removal rate is respectively 90.24% and 4.10mg/L/h, NH4 +It is dense under-N clearance and optimum condition
The NH spent in the synthetic medium for 300mg/L4 +- N clearances are approached, but removal rate is reduced.
It can be seen from Fig. 7 that access arthrobacterium WZUF01 cultivates 72h in animal farm wastewater after the anaerobic digestion of addition sodium acetate
Afterwards, NO3 --N、NO2 -- N and COD is removed and tended towards stability, and control group does not almost decline equally, with NH4 +- N and TN is removed and thalline
Grow the time tended towards stability identical.Find out with reference to table 10, anaerobism anaerobic digestion of the arthrobacterium to WZUF01 to addition sodium acetate
The NO of animal farm wastewater afterwards3 --N、NO2 -- N and COD clearance is respectively 96.3%, 98.12% and 76.84%.
Livestock and poultry breeding industry pollutant emission standard (GB18596-2001) provides that intensive livestock and poultry breeding industry pollution highest is held
Perhaps average daily concentration of emission：Detesting for sodium acetate is added in COD 400mg/L, ammonia nitrogen 80mg/L, the arthrobacterium WZUF01 of the table of comparisons 10 processing
TN, NO after oxygen digestion after animal farm wastewater3 --N、NO2 --N、NH4 +- N and COD concentration, have met livestock and poultry breeding industry pollutant row
Put standard.
Embodiment described above is only the preferred embodiment to absolutely prove the present invention and being lifted, protection model of the invention
Enclose not limited to this.Equivalent substitute or conversion that those skilled in the art are made on the basis of the present invention, in the present invention
Protection domain within.Protection scope of the present invention is defined by claims.
<110>Wenzhou University
<120>One plant of heterotrophic nitrification-aerobic denitrification arthrobacterium and its application
<130> 1
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1594
<212> DNA
<213>Arthrobacterium（Arthrobacter sp.）WZUF01
<400> 1
1 TGCTTACACA TGCAAGTCGA ACGATGAAGC TAAAGCTTGC TTTGGTGGAT
51 TAGTGGCGAA CGGGTGAGTA ACACGTGAGT AACCTGCCCC CGACTCTGGG
101 ATAAGCCTGG GAAACTGGGT CTAATACCGG ATATGACCTC CTACCGCATG
151 GTGGGTGGTG GAAAGATTTA TCGGTGGGGG ATGGACTCGC GGCCTATCAG
201 CTTGTTGGTG AGGTAATGGC TCACCAAGGC GACGACGGGT AGCCGGCCTG
251 AGAGGGTGAC CGGCCACACT GGGACTGAGA CACGGCCCAG ACTCCTACGG
301 GAGGCAGCAG TGGGGAATAT TGCACAATGG GCGAAAGCCT GATGCAGCGA
351 CGCCGCGTGA GGGATGACGG CCTTCGGGTT GTAAACCTCT TTCAGTAGGG
401 AAGAAGCGAA AGTGACGGTA CCTGCAGAAG AAGCGCCGGC TAACTACGTG
451 CCAGCAGCCG CGGTAATACG TAGGGCGCAA GCGTTATCCG GATTTATTGG
501 GCGTAAAGAG CTCGTAGGCG GTTTGTCGCG TCTGCCGTGA AAGTCCGAGG
551 CTCAACCTCG GATCTGCGGT GGGTACGGGC AGACTAGAGT GATGTAGGGG
601 AGACTGGAAT TCCTGGTGTA GCGGTGAAAT GCGCAGATAT CAGGAGGAAC
651 ACCGATGGCG AAGGCAGGTC TCTGGGCATT TACTGACGCT GAGGAGCGAA
701 AGCATGGGGA GCGAACAGGA TTAGATACCC TGGTAGTCCA TGCCGTAAAC
751 GTTGGGCACT AGGTGTGGGG GACATTCCAC GTTTTCCGCG CCGTAGCTAA
801 CGCATTAAGT GCCCCGCCTG GGGAGTACGG CCGCAAGGCT AAAACTCAAA
851 GGAATTGACG GGGGCCCGCA CAAGCGGCGG AGCATGCGGA TTAATTCGAT
901 GCAACGCGAA GAACCTTACC AAGGCTTGAC ATGTTCCAGA CCGGGCTAGA
951 GATAGTCCTT CCCTTCGGGG CTGGTTCACA GGTGGTGCAT GGTTGTCGTC
1001 AGCTCGTGTC GTGAGATGTT GGGTTAAGTC CCGCAACGAG CGCAACCCTC
1051 GTTCCATGTT GCCAGCACGT AGTGGTGGGG ACTCATGGGA GACTGCCGGG
1101 GTCAACTCGG AGGAAGGTGG GGATGACGTC AAATCATCAT GCCCCTTATG
1151 TCTTGGGCTT CACGCATGCT ACAATGGCCG GTACAATGGG TTGCGATACT
1201 GTGAGGTGGA GCTAATCCCT AAAAGCCGGT CTCAGTTCGG ATTGGGGTCT
1251 GCAACTCGAC CCCATGAAGT CGGAGTCGCT AGTAATCGCA GATCAGCAAC
1301 GCTGCGGTGA ATACGTTCCC GGGCCTTGTA CACACCGCCC GTCAAGTCAC
1351 GAAAGTTGGT AACACCCGAA GCCGATGGCC TAACCACCTT GTGT
Claims (9)
1. the arthrobacterium of one plant of heterotrophic nitrification-aerobic denitrification, it is characterised in that the bacterial strain is arthrobacterium（Arthrobactersp.）
WZUF01, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and collection numbering of registering on the books is
CGMCC NO.14012。
2. the application of the arthrobacterium of the heterotrophic nitrification-aerobic denitrification described in claim 1, it is characterised in that the arthrobacterium
WZUF01 is used to go to denitrogenate the nitrogen of sewage：Arthrobacterium WZUF01 is inoculated in nitrogen sewage with except denitrification, the nitrogen sewage be containing
There is NH4 +、NO3 -、NO2 -The sewage of one of them or its combination.
3. application according to claim 2, it is characterised in that the arthrobacterium WZUF01 is used to go to denitrogenate the nitrogen of sewage
Method, comprises the following steps：
1）Preservation strain arthrobacterium WZUF01 is inoculated in activation medium, is cultivated, and centrifugation obtains thalline, and thalline is with sterile washing
OD is made after washing600For 0.9~1.0 bacteria suspension；
2）By step 1）Gained bacterial suspension inoculation is cultivated to denitrogenate in nitrogen sewage；
Wherein, the nitrogen sewage is to contain NH4 +、NO3 -And NO2 -One kind or its combination sewage.
4. application according to claim 3, it is characterised in that step 1）The activation medium is constituted：Peptone
10g, dusty yeast 5g, NaCl 5g, H2O 1000 mL、pH 7.0~7.2；
Step 1）Incubation time be 10 ~ 14h, temperature be 25 ~ 30 DEG C, under the r/min of rotating speed 150 ~ 160 cultivate.
5. application according to claim 4, it is characterised in that step 2）For by step 1）Gained bacterial suspension inoculation is dirty in nitrogen
In water, and it is 4 to add the mass ratio of carbon contained in carbon source, carbon source and nitrogen contained in nitrogen sewage:1~10:1.
6. application according to claim 5, it is characterised in that the carbon source is sodium citrate, sodium acetate, sodium lactate or fourth
The one of which of diacid sodium or its any combination.
7. the application described in claim 3, it is characterised in that step 2）In to go to denitrogenate the cultivation temperature of the nitrogen of sewage be 15 ~ 40
℃。
8. the application described in claim 3, it is characterised in that step 2）In to go to denitrogenate the cultivation temperature of the nitrogen of sewage be 20 ~ 35
℃。
9. the application described in any one of claim 3 ~ 7, it is characterised in that step（2）It is middle by step 1）Gained bacterial suspension inoculation
In nitrogen sewage, pH5 ~ 9 are adjusted.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110184223A (en) * | 2019-06-17 | 2019-08-30 | 华中农业大学 | A kind of Kayseri arthrobacterium that can efficiently remove ammonia nitrogen in aquaculture wastewater and its application |
| CN110964713A (en) * | 2019-12-20 | 2020-04-07 | 武汉理工大学 | Preparation method of immobilized microorganism particles for removing ammonia nitrogen from black and odorous water |
| CN112226388A (en) * | 2020-10-23 | 2021-01-15 | 微米环创生物科技(北京)有限公司 | Novel enzyme-producing species of propionibacteriaceae and application thereof |
| CN112760260A (en) * | 2021-01-26 | 2021-05-07 | 武汉市农业科学院 | Sporobacillus partialis and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7601346B1 (en) * | 2005-12-28 | 2009-10-13 | The United States Of America, As Represented By The Secretary Of Agriculture | Choline-utilizing microbial strains for biologically controlling fusarium head blight |
| CN104862260A (en) * | 2015-06-10 | 2015-08-26 | 国家海洋局第三海洋研究所 | Arthrobacter with aerobic denitrification capability and application thereof |
| CN105586294A (en) * | 2016-01-07 | 2016-05-18 | 温州大学 | Acinetobacter and application of acinetobacter in removal of nitrogen and phosphorus from wastewater |
| CN106399191A (en) * | 2016-10-31 | 2017-02-15 | 杭州朗境环保科技有限公司 | Strain of Arthrobacter sp. B2 and application thereof to nitrogen-containing sewage degradation |
-
2017
- 2017-06-22 CN CN201710480489.0A patent/CN107245463B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7601346B1 (en) * | 2005-12-28 | 2009-10-13 | The United States Of America, As Represented By The Secretary Of Agriculture | Choline-utilizing microbial strains for biologically controlling fusarium head blight |
| CN104862260A (en) * | 2015-06-10 | 2015-08-26 | 国家海洋局第三海洋研究所 | Arthrobacter with aerobic denitrification capability and application thereof |
| CN105586294A (en) * | 2016-01-07 | 2016-05-18 | 温州大学 | Acinetobacter and application of acinetobacter in removal of nitrogen and phosphorus from wastewater |
| CN106399191A (en) * | 2016-10-31 | 2017-02-15 | 杭州朗境环保科技有限公司 | Strain of Arthrobacter sp. B2 and application thereof to nitrogen-containing sewage degradation |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110184223A (en) * | 2019-06-17 | 2019-08-30 | 华中农业大学 | A kind of Kayseri arthrobacterium that can efficiently remove ammonia nitrogen in aquaculture wastewater and its application |
| CN110184223B (en) * | 2019-06-17 | 2020-11-24 | 华中农业大学 | Arthrobacter calsium capable of removing ammonia nitrogen in culture sewage and application thereof |
| CN110964713A (en) * | 2019-12-20 | 2020-04-07 | 武汉理工大学 | Preparation method of immobilized microorganism particles for removing ammonia nitrogen from black and odorous water |
| CN112226388A (en) * | 2020-10-23 | 2021-01-15 | 微米环创生物科技(北京)有限公司 | Novel enzyme-producing species of propionibacteriaceae and application thereof |
| CN112760260A (en) * | 2021-01-26 | 2021-05-07 | 武汉市农业科学院 | Sporobacillus partialis and application thereof |
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