CN109439568A - One plant of denitrifying bacteria and its application and microbial bacterial agent - Google Patents

One plant of denitrifying bacteria and its application and microbial bacterial agent Download PDF

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CN109439568A
CN109439568A CN201811200369.1A CN201811200369A CN109439568A CN 109439568 A CN109439568 A CN 109439568A CN 201811200369 A CN201811200369 A CN 201811200369A CN 109439568 A CN109439568 A CN 109439568A
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bacterial strain
denitrifying bacteria
bacterial agent
culture
microbial
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CN109439568B (en
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杜莉莉
包海花
王明中
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Zhongyuan Environmental Protection Co ltd
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    • C12N1/00Microorganisms, 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
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    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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    • C02F3/34Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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    • C12N1/00Microorganisms, 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
    • C12N1/20Bacteria; Culture media therefor
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00Nature of the contaminant
    • C02F2101/10Inorganic compounds
    • C02F2101/16Nitrogen compounds, e.g. ammonia
    • C02F2101/163Nitrates

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Abstract

The present invention relates to one plant of denitrifying bacterias, are comamonas (Comamonas sp.) DB-1, are deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCC No.16354.The bacterial strain can carry out denitrification denitrogenation under a variety of different carbon sources, and in high concentration nitrate system, and the starting of DB-1 bacterial strain denitrification process is fast, and conversion is thorough;Secondly C/N ratio when conversion nitrate is lower, and carbon source used is few, saves the additional carbon cost of sewage water denitrification processing;DB-1 bacterial strain denitrification process is not interfered in the presence of ammoniacal nitrogen, thus the range of the sewage of the bacterial strain suitable treatment is wider;It is resistant to high alkalinity environment, there is preferable applicability.

Description

One plant of denitrifying bacteria and its application and microbial bacterial agent
Technical field
The invention belongs to technical field of environmental microorganism, are related to one plant of denitrifying bacteria and its application and microbial bacterial agent.
Background technique
Nitrogen is one of indispensable biological element, has important influence to human survival and development, but due to the mankind Excessively development and utilization nitrogen, causes a series of problem of environmental pollution, threatens the Health and Living of the mankind.Nitrogen it is a large amount of Discharge can cause water hypoxia by nitrification, so that water body is blacked smelly;Measures of Algae in Water Body excessive multiplication can be stimulated and caused Eutrophication;In Nitrogen Cycling other than the nitrogen of molecular state, the accumulation such as nitrate of all nitrogen cycle intermediate products and Nitrite can produce serious influence to the mankind and environment.
The nitrogen content in sewage water body can be controlled by physical-chemical process and biological denitrificaion method at present.Physical chemistry Method denitrogenation technology is applied widely, has a corresponding effect to the nitrogenous effluent of various concentration, but its there is also at high cost, fortune Row is complicated, is also easy to produce the disadvantages of secondary pollution.Bio-denitrification technology preferably overcomes the shortcomings that physical-chemical process, has technique Simply, cost is relatively low, is easy the advantages that promoting, and is one of most economical, most efficient method in current sewage water denitrification processing.The party The core of method is nitration reaction and anti-nitration reaction, since the flora difference for participating in reaction is different with sewage treatment process parameter, It is carried out in the reactor that nitrification and denitrification reaction is isolated at two, or alternating anoxic is caused to become reconciled over time and space It is carried out in the same reactor of oxygen environment.There is also some drawbacks for traditional biological denitrification process: first is that common denitrification flora increases It is slow to grow speed, is difficult that high concentration is maintained to carry out denitrogenation, second is that since nitrification process generates acidity, so that pH value of sewage water drops It is low, thus have an adverse effect to anti-nitration reaction;Third is that when the low C/N of process is than sewage, needing to add a large amount of carbon containing has Machine object guarantees denitrification effect, increases cost;Fourth is that conventional denitrification reaction is not thorough, intermediate product NO, the N generated2O etc. Gas long-term accumulation aggravates destruction of the greenhouse effects to environment.
Summary of the invention
In view of this, the purpose of the present invention is to provide one plant of denitrifying bacteria and its applications, and by the denitrifying bacteria The microbial bacterial agent of preparation.
In order to achieve the above objectives, the invention provides the following technical scheme:
1. one plant of denitrifying bacteria, the bacterial strain is comamonas (Comamonas sp.) DB-1, and it is micro- to be deposited in China Biological inoculum preservation administration committee common micro-organisms center, deposit number are CGMCC No.16354.
Denitrifying bacteria DB-1 16SrRNA gene order of the invention is as shown in SEQ ID No.1.
Denitrifying bacteria of the invention, bacterium colony are round, edge clear, and colony colour is white, are moistened, bright, bacterium colony Back side non-pigment, bacterium colony easy picking on plate;It is rod-shaped, Gram-negative bacteria.
2. providing application of the denitrifying bacteria DB-1 in sewage treatment.
Further, the application is to slough the nitrate nitrogen in sewage using denitrifying bacteria DB-1 bacterial strain.
Further, the clump count of denitrifying bacteria is 4.0 × 10 in the processing5-4.0×107cfu/ml。
3. the microbial bacterial agent containing denitrifying bacteria DB-1, the microbial bacterial agent the preparation method comprises the following steps: by bacterial strain DB- 1 seed liquor is seeded to enrichment culture in culture solution by 0.5%-2%v/v, and sterile air is passed through into tank in fermentation process, Mixing speed 180-250rpm, fermentation temperature are 28-35 DEG C, fermentation time 35-45h, collect culture solution, that is, denitrifying bacteria DB- 1 microbial bacterial agent.
Further, the microbial bacterial agent the preparation method comprises the following steps: the seed liquor of bacterial strain DB-1 is seeded to training by 1%v/v Enrichment culture in nutrient solution is passed through sterile air, mixing speed 200rpm in fermentation process into tank, and fermentation temperature is 30 DEG C, Fermentation time 40h collects culture solution, that is, denitrifying bacteria DB-1 microbial bacterial agent.
The beneficial effects of the present invention are: the present invention to pass through the long-term anaerobic acclimation of high nitrogen environment, obtain one plant it is novel anti- Nitrobacteria DB-1 bacterial strain, is sequenced by 16s rDNA, is proved with ncbi database comparison result, this is a new strains, is had More practicabilities.It proves that DB-1 bacterial strain has the performance of ideal degradation total nitrogen through a large number of experiments, shows: 1) in height In concentration nitric acid salt system, the starting of DB-1 bacterial strain denitrification process is fast, and conversion is thorough, and situation that can be lower in bacteria concentration Lower completion conversion, this, which means that, has lower microbial inoculum preparation cost when industry uses;2) C/N ratio when its conversion nitrate Lower, carbon source used is few, to save the additional carbon cost of sewage water denitrification processing;3) its anti-nitre is not interfered in the presence of ammoniacal nitrogen Change process, thus the range of the sewage of the bacterial strain suitable treatment is wider;4) DB-1 bacterial strain denitrification is resistant to the alkalinity of pH8.0 Environment has preferable applicability;5) DB-1 bacterial strain denitrification process is thorough, without intermediate pernicious gas such as NO, N2O etc. is produced Raw, environmental pollution is small.
Detailed description of the invention
In order to keep the purpose of the present invention, technical scheme and beneficial effects clearer, the present invention provides following attached drawing and carries out Illustrate:
Fig. 1 is the 16S rDNA sequence alignment analysis result of bacterial strain DB-1;
Fig. 2 is the flat-plate bacterial colony figure for selecting bacterial strain DB-1.
Biomaterial preservation
Bacterial strain DB-1 in the present invention was deposited in China Committee for Culture Collection of Microorganisms on August 29th, 2018 Common micro-organisms center (abbreviation CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3), deposit number CGMCC No.16354, classification naming are comamonas (Comamonas sp.).
Specific embodiment
Below in conjunction with attached drawing, a preferred embodiment of the present invention will be described in detail.It is not specified in embodiment specific The experimental method of condition, usually according to conventional conditions or according to the manufacturer's recommendations.
DB strain isolation culture medium:
1, liquid denitrification culture medium prescription (g/L): anhydrous sodium acetate: 2g;Dipotassium hydrogen phosphate: 0.4g;Magnesium sulfate: 0.6g;Iron chloride: 0.5g;Calcium chloride: 0.1g;Dipotassium hydrogen phosphate: 0.2g;Sodium nitrate: 1g;Microelement: 2ml;PH=7.0- 7.2。
2, solid denitrification differential medium (g/L): 1%BTB:10g;Anhydrous sodium acetate: 2g;Dipotassium hydrogen phosphate: 0.4g; Magnesium sulfate: 0.6g;Iron chloride: 0.5g;Calcium chloride: 0.1g;Dipotassium hydrogen phosphate: 0.2g;Sodium nitrate: 1g;Microelement: 2ml;pH =7.0-7.2;Agar: 2%.
3, slant preservation culture medium prescription (g/L): beef extract: 1g;Peptone: 10g;Sodium chloride: 5g;PH=7.2-7.4; Agar: 2%;Sterilising conditions: 121 DEG C, 30min.
1 bacterial strain screening of embodiment
DB-1 strain isolation process: taking certain sewage plant sediment of pond mixed liquor 1ml, is connected to the progress of liquid denitrification culture medium The high long-term anaerobic acclimation of nitrogen environment, adds paraffin sealing after being inoculated in culture medium, keeps anaerobic environment, initial screening with air exclusion Test is growth 6-8 days, and the attribute testing in later period is domestication culture 3-4 days in seed culture fluid, grows to OD600=1 or so Amount.After domestication, 10 times of gradient dilutions are carried out with sterile water, take 10-4、10-5、10-6It is coated on solid denitrification and identifies culture On base, each dilution 3 parallel, until there is clear bacterium colony on plate, observes bacterium colony size and color, picking colony is big and all While having the single colonie of blue halos, as shown in Fig. 2, that wherein iris out is No. 3 bacterial strain DB-1 of the invention.It is inoculated in inclined-plane training Base is supported, 4 DEG C of Storage in refrigerator screen altogether 11 plants of bacterial strains, number 1-11.It is anti-that the bacterial strain that primary dcreening operation obtains is inoculated with liquid respectively again Culture medium is nitrified, static gas wave refrigerator 3 days, the liquid medium after 10ml sterilizing is seeded to 5% inoculum concentration, is given birth under room temperature It is 6 days long, measure each culture tube nitrate nitrogen index.The comparison of experimental result is carried out, as shown in table 1.As can be seen that being obtained in primary dcreening operation To 11 plants of bacterial strains in, wherein No. 3, No. 8 and No. 11 bacterial strains convert completely, and its denitrification effect has consistent level.It will This three plants of bacterial strains are identified, wherein No. 8 bacterial strains (being named as DB-3) are identified as pseudomonad, sequence such as SEQ ID It is traditional denitrifying bacterium, using this bacterial strain as reference shown in No.3.
1 primary dcreening operation bacterial strain of table, 6 days degradation nitrate nitrogen index results
Chinese microorganism strain preservation is deposited in by select above No. 3 Strain Designation DB-1, and on August 29th, 2018 Administration committee's common micro-organisms center (abbreviation CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3), deposit number For CGMCC No.16354, classification naming is comamonas (Comamonas sp.).
By select above No. 11 Strain Designation DB-2, and Chinese microorganism strain guarantor was deposited on August 29th, 2018 It hides administration committee's common micro-organisms center (abbreviation CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3), preservation Number be CGMCC No.16355, classification naming be Stenotrophomonas (Stenotrophomonas sp.).
Embodiment 2
The sequencing of bacterial strain DB-1 and taxonomic identification:
Extract 16s rDNA and carry out PCR amplification: 16s rDNA carries out PCR amplification using primer 2 7F/1492R;PCR product Purifying: ExoSApP-IT purifying is carried out to single band PCR product, the PCR product for having non-specific item is carried out to cut glue purification; Sanger PCR sequencing PCR, bidirectional sequencing.DB-1 bidirectional sequencing splice sequence, sequence as shown in SEQ ID No.1, bacterial strain DB-1's 16S rDNA sequence and comparison analysis result are as shown in Figure 1.As shown in Figure 1, the Comamonas sp. of DB-1 and lane database is same Source similitude is 99%, can only be accredited as same category, and 13.58bp is not identical there are also having in 1% otherness, that is, 1358bp, and has been sent out The 16s rDNA sequence of table bacterial strain is not exactly the same, therefore is accredited as novel bacterial and preservation and survival.
Bacterial strain DB-1 carries out Morphological Identification by Observation of biological characteristics, as a result as follows:
By DB-1 streak inoculation on specific solid medium, incubated at room temperature 1-2 days, that is, there is round, edge clear Denitrifying bacteria bacterium colony, colony colour is white, is moistened, bright, bacterium colony back side non-pigment, bacterium colony easy picking on plate; DB-1 is rod-shaped, Gram-negative bacteria.
Embodiment 3
The time course of DB-1 bacterial strain conversion nitrate:
DB-1, DB-2 bacterial strain and 5% inoculum concentration of reference strain DB-3 similarity condition are seeded to liquid denitrification culture medium, Every the content of 24 hours sample detection culture solution nitrate, the results are shown in Table 2, as can be seen from Table 2 in high concentration nitric acid In salt nitrogen systems, DB-1 bacterial strain carry out anti-nitration reaction starting speed it is fast, with DB-3 also in the laundering period compared with, first day nitric acid Salt nitrogen degradation 50%;In addition, DB-1 bacterial strain anti-nitration reaction process is fast, third day is all degraded, and DB-3 then the 5th talent All degradation finishes.
Degradation total nitrogen curve determination result in 2. 3 bacterial strain of table 6 days
Embodiment 4
Counting alive microbial in bacterial strain DB-1 conversion culture solution: the culture solution cultivated in embodiment 33 days is subjected to plate viable bacteria meter Number, calculating bacterium number in DB-1 culture is 4.0 × 106cfu/ml。
Embodiment 5
The research of DB-1 nitrogen removal characteristics
1. influence of the ammoniacal nitrogen to bacterial strain DB-1 denitrogenation
In the ammoniacal nitrogen (NH with 1g/L sodium nitrate equivalent4)2SO4In the presence of, respectively by bacterial strain DB-1, DB-2 and DB-3 carries out aerobic oscillation and anaerobism static gas wave refrigerator, denitrogenation the results are shown in Table 3.DB-1 is amphimicrobe as can be seen from the results, ammoniacal nitrogen In the presence of not influencing its anaerobic denitrifying process.
Influence of 3 ammoniacal nitrogen of table to bacterial strain denitrification index
2. respectively testing bacterial strain DB-1 and DB-3 under variety classes carbon source environment, 4 are shown in Table using experimental data. As can be seen from the results, DB-1 bacterial strain can use a greater variety of carbon sources and reach ideal degradation effect.
6 days index degradation results of strain growth under 4 variety classes carbon source environment of table
3. carbon source difference C/N of the same race is than under environment, the data of DB-1 and DB-3 degradation nitrate are shown in Table 5.As can be seen from the results, DB-1 can reach ideal denitrification effect at lower C/N.
6 days index degradation results of strain growth under 5 difference C/N environment of table
4. bacterial strain DB-1 and DB-3 is shown in Table 6 in high pH environment bacterial strain denitrification effect, data.As can be seen from the results, DB-1 can be with Reach ideal denitrification effect under high alkalinity environment.
64 days index degradation results of pH high environment strain growth of table
Embodiment 6
The preparation of DB-1 microbial inoculum: the 10ml seed liquor cultivated three days is seeded to enrichment culture in 1L culture solution, as hair Ferment seed liquor, the fermentation cylinder for fermentation after being seeded to sterilizing, ferment control temperature are 30 DEG C, are passed through in fermentation process into tank Sterile air;Mixing speed 200rpm, incubation time about 40h collect culture solution, that is, denitrifying bacteria DB-1 microbial inoculum.
Embodiment 7
Central Plains environmental protection Kaifeng 5 side of water utilities advanced nitrogen project pilot-plant volume, DB-1 strain is inoculated with 1% inoculum concentration To pilot-plant culture 2 days, start to intake, flow 0.8-1M3/ h, to pilot-plant inflow and outflow water quality from September 4 in 2018 The continuous monitoring 10 days of September 13 days of -2018 years days, data are shown in Table 7.The nitrate nitrogen known to 7 data of table is down to 0.5mg/ L, total nitrogen is in 2mg/L or less.
7 denitrogenation project testing detection data of table
Finally, it is stated that preferred embodiment above is only used to illustrate the technical scheme of the present invention and not to limit it, although logical It crosses above preferred embodiment the present invention is described in detail, however, those skilled in the art should understand that, can be Various changes are made to it in form and in details, without departing from claims of the present invention limited range.
Sequence table
<110>Central Plains Environmental Protection Co.Ltd.
<120>one plants of denitrifying bacteria and its application and microbial bacterial agents
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1358
<212> DNA
<213> Comamonas sp.
<400> 1
gcgcacttcg gatgctgacg agtggcgaac gggtgagtaa tacatcggaa cgtgcctagt 60
agtgggggat aactactcga aagagtggct aataccgcat gagatctacg gatgaaagca 120
ggggatcgca agaccttgtg ctactagagc ggccgatggc agattaggta gttggtggga 180
taaaagctta ccaagccgac gatctgtagc tggtctgaga ggacgatcag ccacactggg 240
actgagacac ggcccagact cctacgggag gcagcagtgg ggaattttgg acaatgggcg 300
caagcctgat ccagcaatgc cgcgtgcagg atgaaggcct tcgggttgta aactgctttt 360
gtacggaacg aaaagccctg ggttaatacc ctggggtcat gacggtaccg taagaataag 420
caccggctaa ctacgtgcca gcagccgcgg taatacgtag ggtgcgagcg ttaatcggaa 480
ttactgggcg taaagcgtgc gcaggcggtt ttgtaagaca gaggtgaaat ccccgggctc 540
aacctgggaa ctgcctttgt gactgcaagg ctagagtacg gcagaggggg atggaattcc 600
gcgtgtagca gtgaaatgcg tagatatgcg gaggaacacc gatggcgaag gcaatcccct 660
gggcctgtac tgacgctcat gcacgaaagc gtggggagca aacaggatta gataccctgg 720
tagtccacgc cctaaacgat gtcaactggt tgttgggtct taactgactc agtaacgaag 780
ctaacgcgtg aagttgaccg cctggggagt acggccgcaa ggttgaaact caaaggaatt 840
gacggggacc cgcacaagcg gtggatgatg tggtttaatt cgatgcaacg cgaaaaacct 900
tacccacctt tgacatgtac ggaatccttt agagatagag gagtgctcga aagagagccg 960
taacacaggt gctgcatggc tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg 1020
caacgagcgc aacccttgcc attagttgct acgaaagggc actctaatgg gactgccggt 1080
gacaaaccgg aggaaggtgg ggatgacgtc aagtcctcat ggcccttata ggtggggcta 1140
cacacgtcat acaatggctg gtacaaaggg ttgccaaccc gcgaggggga gctaatccca 1200
taaagccagt cgtagtccgg atcgcagtct gcaactcgac tgcgtgaagt cggaatcgct 1260
agtaatcgtg gatcagaatg tcacggtgaa tacgttcccg ggtcttgtac accccgcccg 1320
tcacaccatg ggaacgggtc tcgccagaag taggtaga 1358
<210> 2
<211> 1393
<212> DNA
<213> Stenotrophomonas sp.
<400> 2
tgcagtcgac ggtagcacag aggagcttgc tccttgggtg acgagtggcg gacgggtgag 60
gaatgcatcg gaatctactc tttcgtgggg gataacgtag ggaaacttac gctaataccg 120
catacgacct acgggtgaaa gcaggggacc ttctaggcct tgcgcgattg aatgagccga 180
tgtccgatta gctagttggc ggggtaatgg cccaccaagg cgacgatcgg tagctggtct 240
gagaggatga tcagccacac tggaactgag acacggtcca gactcctacg ggaggcagca 300
gtggggaata ttggacaatg ggcgcaagcc tgatccagcc ataccgcgtg ggtgaagaag 360
gccttcgggt tgtaaagccc ttttgttggg aaagaaaagc agccggctaa tacccggttg 420
ttctgacggt acccaaagaa taagcaccgg ctaacttcgt gccagcagcc gcggtaatac 480
gaagggtgca agcgttactc ggaattactg ggcgtaaagc gtgcgtaggt ggttgtttaa 540
gtctgtcgtg aaagccctgg gctcaacctg ggaactgcga tggaaactgg gcgactagag 600
tgtggtagag ggtagcggaa ttcctggtgt agcagtgaaa tgcgtagata tcaggaggaa 660
catccatggc gaaggcagct acctgggcca acactgacac tgaggcacga aagcgtgggg 720
agcaaacagg attagatacc ctggtagtcc acgccctaaa cgatgcgaac tggatgttgg 780
gtgcactttg gcacgcagta tcgaagctaa cgcgttaagt tcgccgcctg gggagtacgg 840
tcgcaagact gaaactcaaa ggaattgacg ggggcccgca caagcggtgg agtatgtggt 900
ttaattcgat gcaacgcgaa gaaccttacc tggccttgac atgctgagaa ctttccagag 960
atggattggt gccttcggga actcagacac aggtgctgca tggctgtcgt cagctcgtgt 1020
cgtgagatgt tgggttaagt cccgcaacga gcgcaaccct tgtccttagt tgccagcacg 1080
taatggtggg aactctaagg agaccgccgg tgacaaaccg gaggaaggtg gggatgacgt 1140
caagtcatca tggcccttac ggccagggct acacacgtac tacaatggtg gggacagagg 1200
gctgcaaact cgcgagagta agccaatccc agaaacccca tctcagtccg gattggagtc 1260
tgcaactcga ctccatgaag tcggaatcgc tagtaatcgc agatcagcac tgctgcggtg 1320
aatacgttcc cgggccttgt acacaccgcc cgtcacacca tgggagtttg ttgcaccaga 1380
agcaggtagc tta 1393
<210> 3
<211> 1391
<212> DNA
<213> Pseudomonas sp.
<400> 3
ccatgcagtc gagcggatga ggggtgcttg cactctgatt cagcggcgga cgggtgagta 60
atgcctaaga atctgcccga tagtggggga caacgtttcg aaaggaacgc taataccgca 120
tacgtcctac gggagaaagt gggggatctt cggacctcac gctatcggat gagcctaggt 180
cggattagct agttggtgag gtaatggctc accaaggcga cgatccgtaa ctggtctgag 240
aggatgatca gtcacactgg aactgagaca cggtccagac tcctacggga ggcagcagtg 300
gggaatattg gacaatgggc gaaagcctga tccagccatg ccgcgtgtgt gaagaaggtc 360
ttcggattgt aaagcacttt aagttgggag gaagggctgt cggctaatac cctgcagttt 420
tgacgttacc aacagaataa gcaccggcta acttcgtgcc agcagccgcg gtaatacgaa 480
gggtgcaagc gttaatcgga attactgggc gtaaagcgcg cgtaggtggt tcagcaagtt 540
ggatgtgaaa gccccgggct caacctggga actgcatcca aaactactga gctagagtac 600
ggtagagggt ggtggaattt cctgtgtagc ggtgaaatgc gtatatatag gaaggaacac 660
cagtggcgaa ggcgaccacc tggactgata ctgacactga ggtgcgaaag cgtggggagc 720
aaacaggatt agataccctg gtagtccacg ccgtaaacga tgtcaactag ctgttgggtt 780
ccttgagaac ttagtagcga agctaacgcg ataagttgac cgcctgggga gtacggccgc 840
aaggttaaaa ctcaaatgaa ttgacggggg cccgcacaag cggtggagca tgtggtttaa 900
ttcgaagcaa cgcgaagaac cttacctggc cttgacatgc tgagaacttt ccagagatgg 960
attggtgcct tcgggaactc agacacaggt gctgcatggc tgtcgtcagc tcgtgtcgtg 1020
agatgttggg ttaagtcccg taacgagcgc aacccttgtc cttagttacc agcacgttat 1080
ggtgggcact ctaaggagac tgccggtgac aaaccggagg aaggtgggga tgacgtcaag 1140
tcatcatggc ccttacggcc agggctacac acgtgctaca atggtcggta cagagggttg 1200
ccaagccgcg aggtggagct aatctcacaa aaccgatcgt agtccggatc gcagtctgca 1260
actcgactgc gtgaagtcgg aatcgctagt aatcgtgaat cagaatgtca cggtgaatac 1320
gttcccgggc cttgtacaca ccgcccgtca caccatggga gtgggttgct ccagaagtag 1380
ctagtctaac c 1391

Claims (8)

1. one plant of denitrifying bacteria, which is characterized in that the bacterial strain is comamonas (Comamonas sp.) DB-1, preservation In China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No.16354.
2. denitrifying bacteria according to claim 1, which is characterized in that the bacteria 16 S rRNA genes sequence such as SEQ Shown in ID No.1.
3. denitrifying bacteria according to claim 1, which is characterized in that the bacterial strain DB-1 colony colour is white, bar Shape, Gram-negative bacteria.
4. application of the denitrifying bacteria described in claim 1 in sewage treatment.
5. application according to claim 4, which is characterized in that its application is the nitrate sloughed in sewage with the bacterial strain Nitrogen.
6. application according to claim 4, which is characterized in that in the processing clump count of denitrifying bacteria be 4.0 × 105-4.0×107cfu/ml。
7. containing the microbial bacterial agent of denitrifying bacteria described in claim 1, which is characterized in that the system of the microbial bacterial agent Preparation Method are as follows: the seed liquor of bacterial strain DB-1 is seeded to enrichment culture in culture solution by 0.5%-2%v/v, in fermentation process to Sterile air, mixing speed 180-250rpm are passed through in tank, fermentation temperature is 28-35 DEG C, fermentation time 35-45h, collects training Nutrient solution, that is, denitrifying bacteria DB-1 microbial bacterial agent.
8. microbial bacterial agent according to claim 7, which is characterized in that the microbial bacterial agent the preparation method comprises the following steps: will The seed liquor of bacterial strain DB-1 is seeded to enrichment culture in culture solution by 1%v/v, and sterile sky is passed through into tank in fermentation process Gas, mixing speed 200rpm, fermentation temperature are 30 DEG C, fermentation time 40h, collect the micro- life of culture solution, that is, denitrifying bacteria DB-1 Object microbial inoculum.
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102220264A (en) * 2011-05-09 2011-10-19 山东大学 Facultatively anaerobic denitrifying bacteria and application thereof in biological denitrification of water body
CN102465101A (en) * 2010-11-04 2012-05-23 中国石油化工股份有限公司 Denitrification bacterium preparation capable of utilizing nitrite to realize denitrification and use thereof
WO2017165244A1 (en) * 2016-03-19 2017-09-28 Kiverdi, Inc. Microorganisms and artificial ecosystems for the production of protein, food, and useful co-products from c1 substrates

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102465101A (en) * 2010-11-04 2012-05-23 中国石油化工股份有限公司 Denitrification bacterium preparation capable of utilizing nitrite to realize denitrification and use thereof
CN102220264A (en) * 2011-05-09 2011-10-19 山东大学 Facultatively anaerobic denitrifying bacteria and application thereof in biological denitrification of water body
WO2017165244A1 (en) * 2016-03-19 2017-09-28 Kiverdi, Inc. Microorganisms and artificial ecosystems for the production of protein, food, and useful co-products from c1 substrates

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