CN101570738B - Agrobacterium with heterotrophic nitrification-aerobic denitrification capability and application thereof in nitrogenous effluent treatment - Google Patents
Agrobacterium with heterotrophic nitrification-aerobic denitrification capability and application thereof in nitrogenous effluent treatment Download PDFInfo
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Abstract
The invention relates to an agrobacterium strain (Agrobacterium sp.) with heterotrophic nitrification-aerobic denitrification capability and an application thereof in nitrogenous effluent treatment. The bacterial strain is characterized in that: 16S rRNA gene contains nucleotide sequence shown in sequence table with a sequence length of 1418bp and a accession number FJ639330 in Genbank. The preservation number thereof is CGMCC No.2962. The agrobacterium of the invention has not only heterotrophic nitrification capability but also aerobic denitrification capability. During nitrogenous effluenttreatment, and one aerobic phase can cause ammonian to change into gaseous product with high denitrification efficiency and easy operation and tremendous economic benefit compared with the traditionalbiological denitrification process.
Description
Technical field
The present invention relates to a strain edaphic bacillus bacterial strain, particularly relate to and a kind ofly under aerobic condition, the ammonia nitrogen in the waste water can be converted into the edaphic bacillus of gaseous product and the application in nitrogenous effluent treatment thereof.
Background technology
The main existence form of nitrogen element in waste water has molecular nitrogen, organic nitrogen, ammonia-state nitrogen, nitrite nitrogen, nitric nitrogen, and part is present in the nitrogen in sulfohydrate and the prussiate.In untreated raw wastewater, organonitrogen and ammonia nitrogen are the main existence forms of nitrogen.Because the carcinogenesis of the toxic action of ammonia nitrogen, nitrate and nitride be to the acidifying and the eutrophication of the ecosystem, river, lake, coastal ocean, how economic, sewage disposal technology that remove the nitrogen in the water has efficiently become the research emphasis and the focus in water pollution control field.
In the multiple denitrogenation method of having used at present, biological denitrificaion is still the main means that denitrogenation of waste water is handled.Traditional denitrogenation of waste water technology is: the organic nitrogen compound in the sewage at first is converted into ammonia nitrogen in biological process, ammonia nitrogen is translated into nitrate and nitrite by autotrophic bacteria in nitrifying process; Nitrate and nitrite reduction are converted into gaseous product and make denitrogenation of waste water under anoxia condition by denitrifying bacteria then.Nitrated is denitrifying prerequisite, but denitrification process just really reaches the purpose that the nitrogenous compound in the waste water removes.Because nitrobacteria has the intensive aerobic, nitrifying process must be aerobic; And promptly being electron acceptor(EA) with oxygen, the classical inverse nitrifier carries out aerobic repiration under aerobic conditions, be electron acceptor(EA) just when having only anaerobic state with nitrate or nitrite, obtain the energy of synthetic cell body, so the classical inverse nitrifier only can just can carry out denitrification under anaerobic environment.Usually nitrification and denitrification is carried out at aerobic zone and oxygen-starved area respectively according to the theoretical biological denitrification process that grows up of traditional biological denitrogenation, form classification nitration denitrification technology, therefore must build nitrification tank and denitrification pond respectively, so just make classification nitration denitrification technology have a lot of weak points: (1) nitrifying bacteria community rate of propagation is slow, and generation time is long, is difficult to keep higher biological concentration; Cause system's hydraulic detention time longer, organic loading is lower, has increased initial cost and working cost; (2) the denitrification denitrogenation system needs anaerobic environment, for obtaining good denitrification effect, must carry out nitrification liquid and reflux, and has increased power consumption and working cost.
At defective in the traditional technology and deficiency, carrying out both at home and abroad the research that addresses these problems always, make great efforts to seek denitrification microorganism novel, better effects if.The biologist discovers that some bacteriums can carry out heterotrophic nitrification to the organic or inorganic nitrogen compound since the eighties in 20th century.Compare with the autotrophic type nitrobacteria, the growth velocity of heterotroph nitrobacteria is fast, the cell yield height.In addition, the research report confirms that some bacterial strains also can carry out denitrification under aerobic condition.In fact, the investigator has isolated many aerobic denitrifying bacterias from soil and active sludge, as ThiosphaeraPantotropha, Diaphorobacter, Comamonas, Paracoccus Denitrifications, AlcaligenesFaecalis and Microvirgula Aerodenitrificans etc.These are found to be the denitrogenation processing that realizes waste water a kind of new thinking are provided.
It is pointed out that people have found that again some bacteriums can carry out heterotrophic nitrification to the organic or inorganic nitrogen compound along with going deep into of studying.Yet, seldom having having the report of heterotrophic nitrification-aerobic denitrification ability bacterium concurrently, the practical application that the special bacterium of this class is applied to the denitrogenation of waste water treating processes is blank especially.Therefore excavation more has the bacterium of this function, and is applied to the treating processes of actual nitrogenous effluent, and the deficiency that remedies in the traditional biological denitrification process is had epoch making significance.
Summary of the invention
The objective of the invention is to overcome the problem that to take anoxic denitrification, aerobic nitrification staging treating in the existing biological denitrification process, a kind of bacterial strain that has the heterotrophic nitrification-aerobic denitrification ability efficiently with spontaneous growth advantage is provided---edaphic bacillus (Agrobacterium sp.), and the application in the processing of nitrogenous effluent.
Bacterial strain provided by the present invention has following feature: under 37 ℃, cultivate 24h on the nutrient agar after, the bacterium colony projection, full edge is smooth, non-pigment; By being negative at microscopically behind the gramstaining, it is shaft-like that thalline is, and size is (1.5～0.3) μ m * (0.6～1.0) μ m, and single paired arrangement does not form gemma; The 16S rRNA gene sequence characteristic of this bacterial strain: its 16S rRNA has as the nucleotide sequence shown in sequence is represented, sequence length is 1418bp, and the accession number in Genbank is FJ639330.
According to its morphological specificity and physiological and biochemical property and the result for retrieval of 16S rRNA gene order in Genbank thereof, identify that this bacterial strain is edaphic bacillus (Agrobacterium sp.).
The edaphic bacillus that the present invention passes through on March 19th, 2008, is preserved in Beijing that " China Committee for Culture Collection of Microorganisms's common micro-organisms " center ", its preserving number are CGMCC No.2962.
The present invention is achieved by following technical proposals:
Edaphic bacillus (Agrobacterium sp.) bacterial strain LAD9 with heterotrophic nitrification-aerobic denitrification capability, it is characterized in that: the 16S rRNA gene of this edaphic bacillus bacterial strain has the nucleotide sequence shown in sequence table, sequence length is 1418bp, accession number in Genbank is FJ639330, and its preserving number is CGMCC No.2962.
Edaphic bacillus as previously discussed (Agrobacterium sp.) bacterial strain LAD9 is characterized in that: under 33-40 ℃, cultivate 16-32h on the nutrient agar after, the bacterium colony projection, full edge is smooth, non-pigment; By being negative at microscopically behind the gramstaining, it is shaft-like that thalline is, and size is (1.5～0.3) μ m * (0.6～1.0) μ m, and single paired arrangement does not form gemma.
Edaphic bacillus as previously discussed (Agrobacterium sp.) bacterial strain LAD9, the grown cell, the cell suspending liquid that it is characterized in that this bacterial strain can be respectively only nitrogen source with the ammonia nitrogen, organism is that carbon source is carried out the heterotrophic nitrification-aerobic denitrification effect, thereby with ammonia nitrogen removal.
Edaphic bacillus as previously discussed (Agrobacterium sp.) bacterial strain LAD9, the grown cell, the bacterial suspension that it is characterized in that this bacterial strain can be nitrogenous source with nitrate nitrogen or nitrite nitrogen respectively, organism is that carbon source is carried out the aerobic denitrification effect, thereby under aerobic condition nitrate nitrogen or nitrite nitrogen is removed.
The application of edaphic bacillus as previously discussed (Agrobacterium sp.) bacterial strain LAD9 in handling nitrogenous effluent, it is characterized in that in nitrogenous effluent, adding edaphic bacillus (Agrobacterium sp.) bacterial strain LAD9, and carry out aeration, keep dissolved oxygen between 2～6mg/L, can realize the removal of nitrogen in the waste water.
Above-described edaphic bacillus (Agrobacterium sp.) is the microbiobacterial agent of activeconstituents preparation.
Edaphic bacillus of the present invention and application thereof have following beneficial effect compared with prior art:
1. the grown cell of edaphic bacillus of the present invention (Agrobacterium sp.), cell suspending liquid can be the only nitrogen source growth with ammonia nitrogen and nitrate nitrogen respectively all, can carry out the heterotrophic nitrification-aerobic denitrification effect under the condition that aerobic exists, be the newfound bacterium with synchronous nitration and denitrification ability of a strain.This specific character of this bacterial strain can only need can realize by aerobic stage the removal of nitrogen with its direct inoculation in nitrogenous effluent; Solve biological denitrificaion in traditional wastewater treatment and need take the problem of anoxic denitrification, aerobic nitrification staging treating; In addition, simplify technical process, saved the cost of equipment and investment, therefore, had huge economic benefit and environmental benefit;
2. this bacterial strain is applicable to the denitrogenation processing of various nitrogenous effluents, has a extensive future, and has good social benefit;
3. edaphic bacillus of the present invention (Agrobacterium sp.) is inoculated in the waste water that contains ammonia nitrogen and organic carbon, can utilize organic carbon to be sole carbon source, ammonia nitrogen is that only nitrogen source carries out metabolism, the clearance of ammonia nitrogen and total nitrogen reaches 100% and 86.54% respectively in the 12h, and ammonia nitrogen degradation speed reaches 7.95mgN/Lh.
4. most ammonia nitrogen is converted into gaseous product by the heterotrophic nitrification-aerobic denitrification effect in the waste water, and the accumulation of nitrite nitrogen and nitrate nitrogen is less;
With this inoculation in the waste water that contains nitrate nitrogen, can utilize nitrate to carry out metabolism for only nitrogen source, the clearance of nitrate nitrogen reaches 100% in the 28h, and degradation rate is 5.44mgN/Lh, illustrates that bacterial strain has stronger aerobic denitrification ability.This specific character strengthens the practicality of this bacterial strain greatly.
Description of drawings
Accompanying drawing 1 is the microphotograph of edaphic bacillus (Agrobacterium sp.);
Accompanying drawing 2 is the degradation curve of edaphic bacillus (Agrobacterium sp.) to ammonia nitrogen;
Accompanying drawing 3 is the degradation curve of edaphic bacillus (Agrobacterium sp.) to nitrate nitrogen.
In Fig. 2 and Fig. 3:
X-coordinate is: with hour time of expression;
Left side ordinate zou is: the concentration of representing with mg/l;
Right ordinate zou is: OD
600Be the thalline optical density(OD);
NH
4 +-N is an ammonia nitrogen;
TN is a total nitrogen.
Embodiment
Below in conjunction with accompanying drawing and embodiment the present invention is described in further detail:
Embodiment 1
Edaphic bacillus (Agrobacterium sp.) bacterial strain LAD9 with heterotrophic nitrification-aerobic denitrification capability, it is characterized in that: the 16S rRNA gene of this edaphic bacillus bacterial strain has the nucleotide sequence shown in sequence table, sequence length is 1418bp, accession number in Genbank is FJ639330, and its preserving number is CGMCC No.2962.
Embodiment 2
Edaphic bacillus as previously discussed (Agrobacterium sp.) bacterial strain LAD9, it is characterized in that: this edaphic bacillus is to take a sample from the immobilization BAF for the treatment of refuse percolate, a strain gram negative bacterium that obtains through domestication, separation and purifying.
Embodiment 3
Edaphic bacillus as previously discussed (Agrobacterium sp.) bacterial strain LAD9 is characterized in that: under 30 ℃, cultivate 2d on the nutrient agar after, the bacterium colony projection, full edge is smooth, non-pigment; By being negative at microscopically behind the gramstaining, it is shaft-like that thalline is, and size is (1.5～0.3) μ m * (0.6～1.0) μ m, and single paired arrangement does not form gemma.
Edaphic bacillus as previously discussed (Agrobacterium sp.) bacterial strain LAD9 is characterized in that: under 40 ℃, cultivate 3d on the nutrient agar after, the bacterium colony projection, full edge is smooth, non-pigment; By being negative at microscopically behind the gramstaining, it is shaft-like that thalline is, and size is (1.5～0.3) μ m * (0.6～1.0) μ m, and single paired arrangement does not form gemma.
Embodiment 5
Have the separation purification method of edaphic bacillus (Agrobacterium sp.) bacterial strain of heterotrophic nitrification-aerobic denitrification capability, this method has following steps:
1) from the percolate composite processing system, takes out the carrier sample;
2) be inoculated in the 500mL Erlenmeyer flask that 100mL enrichment culture liquid is housed: every L contains 0.95gNaNO
3, 0.7g peptone, 0.5g extractum carnis, 0.15g urea, 0.04g NaCl, 0.15g KH
2PO
4, 0.02g KCl, 0.03gMgSO
47H
2O, 0.20g CaCl
22H
2O, pH 7.0-7.5;
3) bind up with gauze bottleneck, in 30 ℃, 140rpm shaking table shaking culture 4d;
4) be the bacterial classification source with this bacterium liquid that shakes in the bottle, take out 10mL and be seeded in the fresh Erlenmeyer flask that 150mL enrichment culture liquid is housed that culture condition is identical, once every the 4d switching;
5) switching 4 times after, nutrient solution is carried out 10 times of gradient dilutions, diluent is uniformly coated on the nutritional medium: every L contains 1.5g KNO
31.0g KH
2PO
40.06g FeSO
47H
2O; 0.2g CaCl
22H
2O; 1.0gMgSO
47H
2O; 8.5g sodium succinate; 18g agar; 7.0,30 ℃ of constant temperature culture 3d of pH;
6) be that about 100 flat board is selected the variform bacterium colony purifying of ruling repeatedly from colony number, and carry out microscopic examination until obtaining single bacterium colony as the primary dcreening operation bacterial classification;
7) the primary dcreening operation bacterial classification that obtains is carried out performance test;
8) isolate the bacterium that a strain has the heterotrophic nitrification-aerobic denitrification ability efficiently thus, called after LAD9.Embodiment 6
Determining of edaphic bacillus (Agrobacterium sp.) denitrogenation condition
The scope of carbon source
Edaphic bacillus (Agrobacterium sp.) LAD9 in the substratum that contains different carbon sources (glucose, anhydrous sodium acetate, sodium succinate, Seignette salt) 30 ℃, the 160rpm shaking table is cultivated 5d, and ammonia nitrogen all is degraded.Illustrate that bacterial strain LAD9 can grow with above-mentioned carbon source.
The scope of temperature
Agrobacterium sp.LAD9 15～40 ℃ all can well-grown, denitrification effect the best between 30～35 ℃.
The scope of pH
Agrobacterium sp.LAD9 can both grow denitrification effect the best under the condition of meta-alkalescence under 5～12 the condition.
Embodiment 7
Above-described edaphic bacillus (Agrobacterium sp.) is the microbiobacterial agent of activeconstituents preparation.
The evaluation of edaphic bacillus (Agrobacterium sp.)
Edaphic bacillus (Agrobacterium sp.) is to take a sample from the immobilization BAF for the treatment of refuse percolate, a strain gram negative bacterium that obtains through domestication, separation and purifying.
Its physiological and biochemical property is as follows: catalase, oxydase, urease reaction result are positive; Can not carry out Mierocrystalline cellulose and starch hydrolysis reaction; MR result is negative, and VP result is positive; Can utilize propionic salt and under the salinity condition of 2%NaCl, can grow.
The centrifugal column type DNA extraction of the 3S test kit that utilizes Shen, Shanghai energy betting office to produce extracts the genomic dna of bacterial strain LAD9.Genomic dna with extraction is a template, adopts primer 2 7F and 1492R to carry out pcr amplification, and amplified production is delivered the order-checking of Shanghai biotechnology company limited, and sequencing result is seen shown in the sequence table.
Utilize BLAST software that sequencing result is analysed and compared, identify that finally bacterial strain LAD9 is edaphic bacillus (Agrobacterium sp.).
Embodiment 9
The heterotrophic nitrification-aerobic denitrification capability of edaphic bacillus (Agrobacterium sp.) bacterial strain is measured
Be nitrogenous source with the ammonia nitrogen, sodium succinate is under the condition of organic carbon source, implements edaphic bacillus (Agrobacterium sp.) heterotrophic nitrification of ammonia nitrogen is removed and synchronous nitration and denitrification ability mensuration.Concrete implementation step is as follows:
Edaphic bacillus (Agrobacterium sp.) is inoculated in 1L contains 0.5g KNO
3With 0.35g NH
4In the LB substratum of Cl (every liter contains NaCl 5g, Tryptones 10g, yeast extract 5g), prevent the intrusion of assorted bacterium and the growth vigor of maintenance thalline, carry out enrichment culture.The bacterium liquid that cultivation obtains is centrifugal, and optical density(OD) OD is made in the NaCl washing with 0.5% three times
600Bacteria suspension for 1-2.
Get the prepared edaphic bacillus of 10mL (Agrobacterium sp.) bacteria suspension, add three and contain the 90ml test media (every L contains 0.25g NH
4Cl, 1.0g KH
2PO
4, 0.06g FeSO
47H
2O, 0.2g CaCl
22H
2O, 1.0g MgSO
47H
2O, pH 7.0～7.3), seal with 9 layers of gauze, 30 ℃, shaking culture in the shaking table of 180rpm.The substratum of not inoculating bacteria suspension carries out experiment under the equal conditions as blank.Every 2 hours reaction solutions that take a morsel, wherein a part was directly used in and measures the thalline optical density(OD), and rest part is centrifugal 10min under 4000rpm, gets the concentration that supernatant liquor is measured various nitrogenous compounds.
Experimental result is seen accompanying drawing 2.By accompanying drawing 2 as seen, bacterial strain edaphic bacillus (Agrobacterium sp.) still has stronger degradation capability to ammonia nitrogen, heterotrophic nitrification speed height.In initial 8h, ammonia-N removal rate promptly reaches 100%, and degradation rate is 7.95mgN/Lh.From the growth curve of bacterial strain, bacterial strain is in stationary phase in initial 2h, and 2-6h bacterial strain subsequently enters logarithmic phase, and the degraded of ammonia nitrogen is very fast, and its degradation curve almost linearly descends; After this, microorganism growth enters the endogenous respiration phase, and increasing does not almost appear in thalline.
In reaction 12h, the clearance of total nitrogen also reaches 86.54%.Most ammonia nitrogen is removed by synchronous heterotrophic nitrification and aerobic denitrification, and residual nitrogen mainly exists with the form of nitrate nitrogen.Its possible degradation pathway is that ammonia nitrogen is converted into nitrate nitrogen and then nitrate nitrogen is reduced to gaseous product, and different with traditional theory is that these two processes are independently finished by same strain bacterial strain.
This shows that bacterial strain edaphic bacillus (Agrobacterium sp.) has stronger heterotrophic nitrification-aerobic denitrification ability.
Embodiment 10
The aerobic denitrification capability of edaphic bacillus (Agrobacterium sp.) bacterial strain is measured
Be nitrogenous source with the nitrate nitrogen, sodium succinate is under the condition of organic carbon source, implements edaphic bacillus (Agrobacteriumsp.) biological removal to nitrate nitrogen under aerobic condition.Concrete implementation step is as follows:
Get 10mL according to the prepared edaphic bacillus of the method for example 2 (Agrobacterium sp.) bacteria suspension, add three and contain the 90ml test media (every L contains 1.0g KNO
3, 1.0g KH
2PO
4, 0.06g FeSO
47H
2O, 0.2g CaCl
22H
2O, 1.0g MgSO
47H
2O, pH 7.0～7.3), seal with 9 layers of gauze, 30 ℃, shaking culture in the shaking table of 180rpm.The substratum of not inoculating bacteria suspension carries out experiment under the equal conditions as blank.Every 2 hours reaction solutions that take a morsel, wherein a part was directly used in and measures the thalline optical density(OD), and rest part is centrifugal 10min under 4000rpm, gets the concentration that supernatant liquor is measured various nitrogenous compounds.
Experimental result is seen accompanying drawing 3.By accompanying drawing 3 as seen, bacterial strain edaphic bacillus (Agrobacterium sp.) still has stronger removal ability to nitrate nitrogen under aerobic condition, realized the removal to nitrate nitrogen 100% in the 28h, and removal speed is 5.44mgN/Lh; The clearance of total nitrogen also reaches 88.07%.From growth curve of bacteria, in preceding 4h, microorganism is in lag phase, and the speed of growth is slow, almost not to the degraded of nitrate; After this microorganism enters logarithmic phase, and the concentration of nitrate nitrogen almost linearly descends; Behind 24h, 100% nitrate nitrogen is degraded greatly, and microorganism enters endogenous metabolism, and total nitrogen concentration does not have to change substantially.
Embodiment 11
Edaphic bacillus (Agrobacterium sp.) is handled nitrogenous effluent
Add edaphic bacillus (Agrobacterium sp.) in water sample, content is 10
12Individual bacteria/milliliters waste water, pH are 7, ammonia-nitrogen content 120mg/L, and the COD value is 500mg/L, and the 180rpm shaking table is cultivated 40h, and ammonia-N removal rate is more than 90～95%.
Should be noted that; above embodiment is just for the present invention is described in further detail; be not used for the present invention is limited; in the scope that does not break away from design of the present invention and spirit; those of ordinary skills; can carry out various improvement or variation, still belong to the protection domain of the appended claim of the present invention.
Sequence table
<160>1
<210>1
<211>1418
<212>DNA
＜213〉edaphic bacillus (Agrobacterium sp.)
<400>1
1 TGGGGGCGGC?TTACACATGC?AGTCGACGCC?CCGCAAGGGG?AGTGGCAGAC
51 GGGTGAGTAA?CGCGTGGGAA?CATACCCTTT?CCTGCGGAAT?AGCTCCGGGA
101?AACTGGAATT?AATACCGCAT?ACGCCCTACG?GGGGAAAGAT?TTATCGGGGA
151?AGGATTGGCC?CGCGTTGGAT?TAGCTAGTTG?GTGGGGTAAA?GGCCTACCAA
201?GGCGACGATC?CATAGCTGGT?CTGAGAGGAT?GATCAGCCAC?ATTGGGACTG
251?AGACACGGCC?CAAACTCCTA?CGGGAGGCAG?CAGTGGGGAA?TATTGGACAA
301?TGGGCGCAAG?CCTGATCCAG?CCATGCCGCG?TGAGTGATGA?AGGCCTTAGG
351?GTTGTAAAGC?TCTTTCACCG?GAGAAGATAA?TGACGGTATC?CGGAGAAGAA
401?GCCCCGGCTA?ACTTCGTGCC?AGCAGCCGCG?GTAATACGAA?GGGGGCTAGC
451?GTTGTTCGGA?ATTACTGGGC?GTAAAGCGCA?CGTAGGCGGA?TATTTAAGTC
501?AGGGGTGAAA?TCCCAGAGCT?CAACTCTGGA?ACTGCCTTTG?ATACTGGGTA
551?TCTTGAGTAT?GGAAGAGGTA?AGTGGAATTC?CGAGTGTAGA?GGTGAAATTC
601?GTAGATATTC?GGAGGAACAC?CAGTGGCGAA?GGCGGCTTAC?TGGTCCATTA
651?CTGACGCTGA?GGTGCGAAAG?CGTGGGGAGC?AAACAGGATT?AGATACCCTG
701?GTAGTCCACG?CCGTAAACGA?TGAATGTTAG?CCGTCGGGCA?GTATACTGTT
751?CGGTGGCGCA?GCTAACGCAT?TAAACATTCC?GCCTGGGGAG?TACGGTCGCA
801?AGATTAAAAC?TCAAAGGAAT?TGACGGGGGC?CCGCACAAGC?GGTGGAGCAT
851?GTGGTTTAAT?TCGAAGCAAC?GCGCAGAACC?TTACCAGCTC?TTGACATTCG
901?GGGTTTGGGC?AGTGGAGACA?TTGTCCTTCA?GTTAGGCTGG?CCCCAGAACA
951?GGTGCTGCAT?GGCTGTCGTC?AGCTCGTGTC?GTGAGATGTT?GGGTTAAGTC
1001CCGCAACGAG?CGCAACCCTC?GCCCTTAGTT?GCCAGCATTT?AGTTGGGCAC
1051TCTAAGGGGA?CTGCCGGTGA?TAAGCCGAGA?GGAAGGTGGG?GATGACGTCA
1101AGTCCTCATG?GCCCTTACGG?GCTGGGCTAC?ACACGTGCTA?CAATGGTGGT
1151GACAGTGGGC?AGCGAGACAG?CGATGTCGAG?CTAATCTCCA?AAAGCCATCT
1201CAGTTCGGAT?TGCACTCTGC?AACTCGAGTG?CATGAAGTTG?GAATCGCTAG
1251TAATCGCAGA?TCAGCATGCT?GCGGTGAATA?CGTTCCCGGG?CCTTGTACAC
1301ACCGCCCGTC?ACACCATGGG?AGTTGGTTTT?ACCCGAAGGT?AGTGCGCTAA
1351CCGCAAGGAG?GCAGCTAACC?ACGGTAGGGT?CAGCGACTGG?GGTGAAGTCG
1401TAACAAGAGC?CATGCCTA
Claims (5)
1. the edaphic bacillus Agrobacterium sp. bacterial strain LAD9 that has heterotrophic nitrification-aerobic denitrification capability, it is characterized in that: this edaphic bacillus bacterial strain is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, preserving number is CGMCC No.2962, the 16SrRNA gene of this edaphic bacillus bacterial strain has the nucleotide sequence shown in SEQID:1, and sequence length is 1418bp.
2. edaphic bacillus Agrobacterium sp. bacterial strain LAD9 as claimed in claim 1, it is characterized in that: this edaphic bacillus is to take a sample from the immobilization BAF for the treatment of refuse percolate, a strain gram negative bacterium that obtains through domestication, separation and purifying.
3. edaphic bacillus Agrobacterium sp. bacterial strain LAD9 according to claim 1 is characterized in that: under 30-40 ℃, cultivate 16-32h on the nutrient agar after, the bacterium colony projection, full edge is smooth, non-pigment; By being negative at microscopically behind the gramstaining, it is shaft-like that thalline is, and size is 1.5～0.3 μ m * 0.6～1.0 μ m, and single paired arrangement does not form gemma.
4. edaphic bacillus Agrobacterium sp. bacterial strain LAD9 according to claim 1, the grown cell, the cell suspending liquid that it is characterized in that this bacterial strain can be only nitrogen source with the ammonia nitrogen all, organism is that carbon source is carried out the heterotrophic nitrification-aerobic denitrification effect, thereby with ammonia nitrogen removal.
5. edaphic bacillus Agrobacterium sp. bacterial strain LAD9 according to claim 1, the grown cell, the bacterial suspension that it is characterized in that this bacterial strain can be nitrogenous source with nitrate nitrogen or nitrite nitrogen all, organism is that carbon source is carried out the aerobic denitrification effect, thereby under aerobic condition nitrate nitrogen or nitrite nitrogen is removed.
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2009
- 2009-04-28 CN CN2009100830143A patent/CN101570738B/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103710294A (en) * | 2014-01-09 | 2014-04-09 | 北京大学 | Strain of efficient ammonifying bacteria and application thereof in organic nitrogen-containing waste water treatment |
| CN103710294B (en) * | 2014-01-09 | 2015-08-12 | 北京大学 | One plant height is imitated Ammonifying bacteria and is being contained the application in organonitrogen wastewater treatment |
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