CN101560487B - Comamonas testosteroni strain for biological denitrificaion and application thereof - Google Patents
Comamonas testosteroni strain for biological denitrificaion and application thereof Download PDFInfo
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- CN101560487B CN101560487B CN2009100851883A CN200910085188A CN101560487B CN 101560487 B CN101560487 B CN 101560487B CN 2009100851883 A CN2009100851883 A CN 2009100851883A CN 200910085188 A CN200910085188 A CN 200910085188A CN 101560487 B CN101560487 B CN 101560487B
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Abstract
The invention discloses a comamonas testosteroni strain for biological denitrificaion and application thereof. The comamonas testosteroni strain GAD4 is preserved in China General Microbiological Culture Collection Center (CGMCC for short) with preservation number CGMCC No. 2963 in March 19th, 2009. The strain is added to nitrogenous effluent with 2 to 6mg/L of dissolved oxygen to realize biological denitrificaion. The strain not only has the capability of heterotrophic nitrification, but also has the capability of aerobic denitrification. In the process of treating the nitrogenous effluent, only one aerobic stage is needed, the denitrification efficiency is high, and the operation is convenient and quick; and compared with the conventional biological denitrification process, the application has huge economic benefits.
Description
Technical field
The present invention relates to the biological denitrificaion field, particularly a kind of Comamonas testosteroni strain and application thereof that is used for biological denitrificaion.
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.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) technology is tediously long.Because nitrification and denitrification effect aerobism difference must be built nitrification tank and denitrification pond respectively, form classification nitration denitrification technology, increased initial cost and working cost; (2) energy consumption is big.Ammonia nitrogen is nitrated to need the oxygen consumption energy supply, and the Prepositive denitrification system must carry out the nitrification liquid internal reflux, has increased power consumption and working cost; (3) denitrifying bacterium will have carbon source as electron donor, if carbon/nitrogen compared lowly in the sewage, then need add organic carbons such as methyl alcohol, and this has not only increased working cost, has also increased the difficulty of operational management and subsequent disposal; (4) nitrifying bacteria community autotrophic bacteria normally, propagation slowly is difficult to keep higher biological concentration, and is eliminated in biological wastewater treatment easily.
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 Thiosphaera Pantotropha, Diaphorobacter, Comamonas, ParacoccusDenitrifications, Alcaligenes Faecalis 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 also not to 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 at present.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 object of the present invention is to provide a kind of Comamonas testosteroni strain and application thereof that is used for biological denitrificaion, this bacterial strain has the heterotrophic nitrification-aerobic denitrification ability concurrently.
Comamonas testosteroni provided by the invention (Comamonas testosteroni) GAD4 is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC) on March 19th, 2009, and preserving number is CGMCC № 2963.
Bacterial strain GAD4 takes a sample from the combined system for the treatment of refuse percolate, a strain gram negative bacterium that obtains through domestication, separation and purifying.
Morphological specificity: bacterial strain GAD4 under 30-40 ℃, cultivate 16-32h on the nutrient agar after, bacterium colony smooth surface, non-pigment; By being negative at microscopically behind the gramstaining, it is shaft-like that thalline is, and size is the μ m of (0.5-1.0) μ m * (1.5-5.0), and the single or paired appearance of cell has end to generate the flagellum of bundle, does not produce gemma.
Physiological and biochemical property: catalase, oxydase, urease reaction result are positive; Can not carry out the cellulose hydrolysis reaction; MR and VP result are negative; Can be that carbon source is grown with the Trisodium Citrate; Can utilize propionic salt; Can not utilize glucose, fructose, semi-lactosi, rhamnosyl, wood sugar, lactose, seminose and maltose fermentation.
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 1462bp.
According to its morphological specificity and physiological and biochemical property and 16S rRNA gene order thereof, identify that this bacterial strain is Comamonas testosteroni (Comamonas testosteroni).
This bacterial strain can be used to denitrogenation, in actual applications, bacterial strain GAD4 can be placed nitrogenous liquid, carries out biological denitrificaion.The dissolved oxygen of described nitrogenous liquid can be 2-6mg/L.
The pH of described nitrogenous liquid can be 6-12, is preferably 7-11.
The temperature of described nitrogenous liquid can be 15-40 ℃, is preferably 30-35 ℃.
Carbon source in the described nitrogenous liquid can be anhydrous sodium acetate, sodium succinate, Seignette salt and/or Trisodium Citrate.
Nitrogen in the described nitrogenous liquid is ammonia nitrogen, nitrate nitrogen and/or nitrite nitrogen.
Another purpose of the present invention is to provide a kind of microbiobacterial agent that is used for biological denitrificaion, and its activeconstituents is described bacterial strain GAD4.
Comamonas testosteroni of the present invention and application thereof have following beneficial effect compared with prior art:
1. the grown cell of bacterial strain GAD4 of the present invention, cell suspending liquid can be the only nitrogen source growth with ammonia nitrogen or nitrate nitrogen respectively all, can carry out the heterotrophic nitrification-aerobic denitrification effect under aerobic conditions, 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. bacterial strain GAD4 of the present invention is inoculated in the waste water that initial ammonia nitrogen concentration is 75mg/L, can utilize organism to be sole carbon source, ammonia nitrogen is that only nitrogen source carries out metabolism, and the clearance of ammonia nitrogen and total nitrogen reaches 90.85% and 84.45% respectively in the 12h, and ammonia nitrogen degradation speed is 5.79mgN/Lh; 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;
4. it is in the waste water of 160mg/L that bacterial strain GAD4 of the present invention is inoculated into initial nitrate nitrogen concentration, can utilize nitrate to carry out metabolism for only nitrogen source, the clearance of nitrate nitrogen reaches 64.05% in the 24h, degradation rate is 4.25mgN/Lh, illustrates that bacterial strain has good aerobic denitrification ability;
5, bacterial strain GAD4 of the present invention is inoculated in the ammonia nitrogen or nitrate nitrogen waste water that starting point concentration is 400mg/L, still has degradation effect and denitrogenation ability preferably, and ammonia nitrogen and nitrate nitrogen degradation rate are respectively 3.14mgN/Lh and 3.12mgN/Lh;
6, bacterial strain GAD4 of the present invention is inoculated in the nitrate nitrogen waste water of different pH values, and the degradation capability of bacterial strain is stronger under the condition of neutrality and meta-alkalescence, and this moment, pH was little to the influence of degradation effect, and this specific character strengthens the practicality of this bacterial strain greatly.
Description of drawings
Fig. 1 is the microphotograph of bacterial strain GAD4;
Fig. 2 is the degradation curve of bacterial strain GAD4 to ammonia nitrogen;
Fig. 3 is the degradation curve of bacterial strain GAD4 to nitrate nitrogen;
Fig. 4 is the degradation curve of bacterial strain GAD4 to ammonia nitrogen;
Fig. 5 is the degradation curve of bacterial strain GAD4 to nitrate nitrogen;
Fig. 6 is bacterial strain GAD4 degraded to ammonia nitrogen under the different rotating speeds condition;
Fig. 7 is bacterial strain GAD4 degraded to nitrate nitrogen under condition of different pH;
Embodiment
The invention will be further described below in conjunction with specific embodiment, but the present invention is not limited to following examples.
Among the following embodiment, if no special instructions, be ordinary method.
Among the following embodiment, described percentage composition is the quality percentage composition if no special instructions.
Embodiment 1. has acquisition and the biological denitrificaion experiment of the bacterial strain GAD4 of heterotrophic nitrification-aerobic denitrification capability
One, the separation purification method that has the bacterial strain GAD4 of heterotrophic nitrification-aerobic denitrification capability
This method has following steps:
1) from Shenzhen Buji refuse landfill percolate composite processing system, takes out the mud sample;
2) be inoculated in the 500mL Erlenmeyer flask that 100mL enrichment culture liquid is housed: every L contains 0.95g NaNO
3, 0.7g peptone, 0.5g extractum carnis, 0.15g urea, 0.04g NaCl, 0.15g KH
2PO
4, 0.02g KCl, 0.03g MgSO
47H
2O, 0.20g CaCl
22H
2O, wherein nutrient solution pH is 7.0-7.5;
3) bind up with gauze bottleneck, in 30 ℃, shaking table shaking culture 4d under the 140rpm condition;
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; Wherein pH in culture medium is 7.0,30 ℃ of constant temperature culture 3d;
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 bacterium-bacterial strain GAD4 that a strain has the heterotrophic nitrification-aerobic denitrification ability efficiently thus.
Two, with ammonia nitrogen (NH
4 +-N) be the biological denitrificaion experiment of nitrogenous source
With the ammonia nitrogen is nitrogenous source, and sodium succinate is an organic carbon source, implements bacterial strain GAD4 the removal ability of ammonia nitrogen is measured.Concrete implementation step is as follows:
Bacterial strain GAD4 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 the NaCl solution washing with 0.05% three times is made optical density(OD) OD
600Bacteria suspension for 1-2.
Get the prepared bacterial strain GAD4 bacteria suspension of 10mL, add that (every L substratum contains 0.29g NH in three culturing bottles that contain the 90ml test media
4Cl, 1.0g KH
2PO
4, 0.06g FeSO
47H
2O, 0.2gCaCl
22H
2O, 1.0g MgSO
47H
2O, 8.5g sodium succinate, pH 7.0-7.3), seal with 9 layers of gauze, 30 ℃, shaking culture in the shaking table of 180rpm (cyclotron radius 15mm).The substratum of not inoculating bacteria suspension carries out experiment under the equal conditions as blank.Extracted reaction solution every 2 hours, wherein a part is directly used in and measures thalline optical density(OD) (OD
600), rest part is centrifugal 10min under 4000rpm, gets the concentration that supernatant liquor is measured various nitrogenous compounds.
The result as shown in Figure 2, bacterial strain GAD4 has stronger degradation capability to ammonia nitrogen, heterotrophic nitrification speed height.In the 12h during reaction, ammonia-N removal rate promptly reaches 90.85%.From the growth curve of bacterial strain, bacterial strain is in stationary phase in initial 2h, and 2-8h 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.
Simultaneously, the clearance of total nitrogen (TN) also reaches 84.45%.Most ammonia nitrogen is removed by the heterotrophic nitrification-aerobic denitrification effect, and residual nitrogen mainly exists with the form of nitrate nitrogen.
The approach of bacterial strain GAD4 degradation of ammonia nitrogen is that ammonia nitrogen is converted into nitrate nitrogen in the step 2, and nitrate nitrogen is reduced to gaseous product then, and different with traditional method is that these two processes are independently finished by same strain bacterial strain.
This shows, be nitrogenous source with the ammonia nitrogen, and bacterial strain GAD4 has stronger heterotrophic nitrification-aerobic denitrification ability.
Three, with nitrate nitrogen (NO
3 --N) be the biological denitrificaion experiment of nitrogenous source
Get the bacteria suspension of 10mL, join that (every L substratum contains 1.16g KNO in three culturing bottles that contain the 90ml test media according to the prepared bacterial strain GAD4 of the method for step 2
3, 1.0g KH
2PO
4, 0.06gFeSO
47H
2O, 0.2g CaCl
22H
2O, 1.0g MgSO
47H
2O, the 8.5g sodium succinate, pH 7.0～7.3), seal with 9 layers of gauze, 30 ℃, shaking culture in the shaking table of 180rpm (cyclotron radius 15mm).The substratum of not inoculating bacteria suspension carries out experiment under the equal conditions as blank.Extracted reaction solution every 2 hours, and under 4000rpm centrifugal 10min, get the concentration that supernatant liquor is measured various nitrogenous compounds.
The result as shown in Figure 3, bacterial strain GAD4 has stronger removal ability to nitrate nitrogen under aerobic condition, realized the removal to nitrate nitrogen 64.05% in the 24h, removing speed is 4.25mg nitrogen/Lh (mgN/Lh); The clearance of total nitrogen is 63.28%, nitrite nitrogen (NO almost do not occur
2 --N) accumulation.In the initial 4-6h, the concentration of nitrate nitrogen remains unchanged substantially, illustrates that microorganism is in stationary phase; After this microorganism enters logarithmic phase, and curve linearly descends.Prove absolutely that bacterial strain can be that electron acceptor(EA) carries out denitrification with nitrate nitrogen and nitrite nitrogen under aerobic condition, the ability of aerobic denitrification is good.
Four, the biological denitrificaion of nitrogenous effluent experiment
Add bacterial strain GAD4 in the waste water (waste water quality is as follows: pH is 7.5, ammonia-nitrogen content 30mg/L, nitrate nitrogen content 6mg/L, COD 550mg/L), make that bacterial concentration is 10 in the water sample
12Individual bacteria/milliliters waste water, dissolved oxygen maintains 4-6mg/L, and temperature is 25-30 ℃, continuous aeration 30h, experiment repeats 3 times, and the result is as shown in table 1, and the COD clearance is 92%, and ammonia-N removal rate is 99%, and nitrogen removal rate is 78%.
The biological denitrificaion result of table 1. waste water
| COD(mg/L) | Ammonia nitrogen (mg/L) | Total nitrogen (mg/L) | |
| Before the processing | 550 | 30 | 36 |
| After the processing | 44±1.23 | 0.3±0.05 | 7.92±1.15 |
| Clearance | 92％±0.22％ | 99％±0.17％ | 78％±3.19％ |
The experiment of embodiment 2. biological denitrificaions
Bacterial strain GAD4 that the method that provides with embodiment 1 prepares and bacteria suspension thereof carry out the biological denitrificaion experiment.
One, with ammonia nitrogen (NH
4 +-N) be the biological denitrificaion experiment of nitrogenous source
Get the prepared bacterial strain GAD4 bacteria suspension of 5mL, add that (every L substratum contains 1.52g NH in three culturing bottles that contain the 95ml test media
4Cl, 1.0g KH
2PO
4, 0.06g FeSO
47H
2O, 0.2gCaCl
22H
2O, 1.0g MgSO
47H
2O, 8.1g Trisodium Citrate, pH are 8.0), seal with 9 layers of gauze, 15 ℃, shaking culture in the shaking table of 160rpm (rotation radius 15mm).The substratum of not inoculating bacteria suspension carries out experiment under the equal conditions as blank.Extract reaction solution every other day, and under 4000rpm centrifugal 10min, get the concentration that supernatant liquor is measured various nitrogenous compounds.
The result when bacterial strain GAD4 is 400mg/L at the ammonia nitrogen starting point concentration, still has stronger degradation capability as shown in Figure 4, heterotrophic nitrification speed height.In first day of inoculation, the degradation rate of ammonia nitrogen is very fast, and clearance reaches 37.75%; The concentration of ammonia nitrogen downward trend linearly in three days afterwards, ammonia-N removal rate is 80% in the final 96h, degradation rate is 3.14mgN/Lh.
Two, with nitrate nitrogen (NO
3 --N) be the biological denitrificaion experiment of nitrogenous source
Get the bacteria suspension of 5mL, join that (every L substratum contains 2.89g KNO in three culturing bottles that contain the 95ml test media according to the prepared bacterial strain GAD4 of the method for embodiment 1
3, 1.0g KH
2PO
4, 0.06gFeSO
47H
2O, 0.2g CaCl
22H
2O, 1.0g MgSO
47H
2O, 8.1g Trisodium Citrate, pH are 8.0), seal with 9 layers of gauze, 15 ℃, shaking culture in the shaking table of 160rpm (rotation radius 25mm).The substratum of not inoculating bacteria suspension carries out experiment under the equal conditions as blank.Extract reaction solution every other day, and under 4000rpm centrifugal 10min, get the concentration that supernatant liquor is measured various nitrogenous compounds.
The result as shown in Figure 5, when bacterial strain GAD4 was 400mg/L at initial nitrate concentration, the clearance of nitrate nitrogen reached 74.91% in the 96h, degradation rate is 3.12mgN/Lh; Total nitrogen concentration is reduced to 115mg/L from initial 400mg/L, and clearance is 71.25%.
A spot of accumulation appearred in nitrite nitrogen in the degradation process, reached peak value 60.71mg/L at the 3rd day.This shows that aerobic denitrifying bacteria is similar with the degradation process of anaerobic denitrifying bacteria, all is that the experience nitrate nitrogen is converted into nitrite nitrogen, nitrite nitrogen and then be converted into two processes of nitrogen.Yet the degradation rate of aerobic denitrifying bacteria will be higher than anaerobic denitrifying bacteria.
The experiment of embodiment 3. biological denitrificaions
Bacterial strain GAD4 that the method that provides with embodiment 1 prepares and bacteria suspension thereof carry out the biological denitrificaion experiment.
One, with ammonia nitrogen (NH
4 +-N) be the biological denitrificaion experiment of nitrogenous source
Get the prepared bacterial strain GAD4 bacteria suspension of 5mL, add that (every L substratum contains 0.42g NH in 4 culturing bottles that contain the 95ml test media
4Cl, 1.0g KH
2PO
4, 0.06g FeSO
47H
2O, 0.2gCaCl
22H
2O, 1.0g MgSO
47H
2O, 7.72g anhydrous sodium acetate, pH are 8.5).Seal with 9 layers of gauze, 40 ℃ is shaking culture in the shaking table of 100rpm, 150rpm and 200rpm (rotation radius 15mm) at rotating speed respectively.The substratum of not inoculating bacteria suspension carries out experiment under the equal conditions as blank.Cultivate 24h and get and shake reaction solution in the bottle, and under 4000rpm centrifugal 10min, get the concentration that supernatant liquor is measured various nitrogenous compounds.
The result as shown in Figure 6, bacterial strain GAD4 is under 100rpm, 150rpm and the 200rpm ammonia-N removal rate to be respectively 64.45%, 89.76% and 84.88% in shaking speed; Clearance to total nitrogen is respectively 61.62%, 83.23% and 75.09%.The dissolved oxygen levels of the difference of shaking speed in can the indirect reaction system, when shaking speed was 100rpm, dissolved oxygen levels was lower in the system, and it is relatively low that ammonia nitrogen and total nitrogen are removed speed; When shaking speed was 200rpm, clearance was lower than the situation that rotating speed is 150rpm slightly, illustrated that hunting speed is excessive and crossed the low growth that all is unfavorable for bacterial classification, and the optimum shaking speed of bacterial strain GAD4 is 150rpm.
Two, with nitrate nitrogen (NO
3 --N) be the biological denitrificaion experiment of nitrogenous source
Get the prepared bacterial strain GAD4 bacteria suspension of 5mL, add that (every L substratum contains 1.0g KNO in 9 culturing bottles that contain the 95ml test media
3, 1.0g KH
2PO
4, 0.06g FeSO
47H
2O, 0.2g CaCl
22H
2O, 1.0g MgSO
47H
2O, the 7.72g anhydrous sodium acetate), initial pH is respectively 4.0,5.0,6.0,7.0,8.0,9.0,10.0,11.0.Seal with 9 layers of gauze, 40 ℃, shaking culture in the shaking table of 160rpm (rotation radius is 15mm).The substratum of not inoculating bacteria suspension carries out experiment under the equal conditions as blank.Cultivate 24h and get and shake reaction solution in the bottle, and under 4000rpm centrifugal 10min, get the concentration that supernatant liquor is measured various nitrogenous compounds.
The result as shown in Figure 7, bacterial strain GAD4 has degradation effect preferably to nitrate nitrogen under the condition of neutral meta-alkalescence, remove poor effect under acidic conditions.When pH greater than 7 the time, the clearance of total nitrogen is 65% in the 24h; When pH greater than 9 the time, nitric efficiency does not have sharply to descend but maintains certain level, illustrates that bacterial strain GAD4 is stronger to the adaptability of alkaline environment.Therefore, the pH of described nitrogenous liquid can be 6-12, is preferably 7-11.This specific character strengthens the practicality of this bacterial strain greatly.
Sequence table
＜110〉Peking University
＜120〉be used for the Comamonas testosteroni strain and the application thereof of biological denitrificaion
<130>CGGNARL92320
<160>1
<210>1
<211>1462
<212>DNA
＜213〉Comamonas testosteroni (Comamobas teststeroni)
<400>1
catgctttaa?catgcaagtc?gaacggtaac?aggtcttcgg?atgctgacga?gtggcgaacg 60
ggtgagtaat?acatcggaac?gtgcctagta?gtgggggata?actactcgaa?agagtagcta 120
ataccgcatg?agatctacgg?atgaaagcag?gggacctttg?ggccttgtgc?tactagagcg 180
gctgatggca?gattaggtag?ttggtggggt?aaaggcttac?caagcctgcg?atctgtagct 240
ggtctgagag?gacgaccagc?cacactggga?ctgagacacg?gcccagactc?ctacgggagg 300
cagcagtggg?gaattttgga?caatgggcga?aagcctgatc?cagcaatgcc?gcgtgcagga 360
tgaaggccct?cgggttgtaa?actgcttttg?tacggaacga?aaagcctggg?gctaatatcc 420
ccgggtcatg?acggtaccgt?aagaataagc?accggctaac?tacgtgccag?cagccgcggt 480
aatacgtagg?gtgcaagcgt?taatcggaat?tactgggcgt?aaagcgtgcg?caggcggttt 540
tgtaagacag?tggtgaaatc?cccgggctca?acctgggaac?tgccattgtg?actgcaaggc 600
tagagtgcgg?cagaggggga?tggaattccg?cgtgtagcag?tgaaatgcgt?agatatgcgg 660
aggaacaccg?atggcgaagg?caatcccctg?ggcctgcact?gacgctcatg?cacgaaagcg 720
tggggagcaa?acaggattag?ataccctggt?agtccacgcc?ctaaacgatg?tcaactggtt 780
gttgggtctt?aactgactca?gtaacgaagc?taacgcgtga?agttgaccgc?ctggggagta 840
cggccgcaag?gttgaaactc?aaaggaattg?acggggaccc?gcacaagcgg?tggatgatgt 900
ggtttaattc?gatgcaacgc?gaaaaacctt?acccaccttt?gacatggcag?gaacttacca 960
gagatggttt?ggtgctcgaa?agagaacctg?cacacaggtg?ctgcatggct?gtcgtcagct 1020
cgtgtcgtga?gatgttgggt?taagtcccgc?aacgagcgca?acccttgcca?ttagttgcta 1080
cattcagttg?agcactctaa?tgggactgcc?ggtgacaaac?cggaggaagg?tggggatgac 1140
gtcaagtcct?catggccctt?ataggtgggg?ctacacacgt?catacaatgg?ctggtacaaa 1200
gggttgccaa?cccgcgaggg?ggagctaatc?ccataaagcc?agtcgtagtc?cggatcgcag 1260
tctgcaactc?gactgcgtga?agtcggaatc?gctagtaatc?gtggatcaga?atgtcacggt 1320
gaatacgttc?ccgggtcttg?tacacaccgc?ccgtcacacc?atgggagcgg?gtctcgccag 1380
aagtaggtag?cctaaccgca?aggagggcgc?ttaccacggc?ggggttcgtg?actggggtga 1440
agtcgtaaca?agagccagac?gg 1462
Claims (9)
1. Comamonas testosteroni (Comamonas testosteroni) bacterial strain GAD4, its preserving number is CGMCC № 2963.
2. the described preserving number of claim 1 is the application of Comamonas testosteroni (Comamonas testosteroni) bacterial strain GAD4 in denitrogenation of CGMCC № 2963.
3. application according to claim 2 is characterized in that: described denitrogenation is carried out in nitrogenous liquid, and the dissolved oxygen of described nitrogenous liquid is 4-6mg/L.
4. according to claim 2 or 3 described application, it is characterized in that: the temperature of described nitrogenous liquid is 15-40 ℃.
5. application according to claim 4 is characterized in that: the temperature of described nitrogenous liquid is 30-35 ℃.
6. application according to claim 2 is characterized in that: the pH of described nitrogenous liquid is 6-12.
7. application according to claim 6 is characterized in that: the pH of described nitrogenous liquid is 7-11.
8. application according to claim 3 is characterized in that: the nitrogen in the described nitrogenous liquid is ammonia nitrogen, nitrate nitrogen and/or nitrite nitrogen.
9. microbiobacterial agent that is used for biological denitrificaion, its activeconstituents is that the described preserving number of claim 1 is Comamonas testosteroni (Comamonas testosteroni) the bacterial strain GAD4 of CGMCC № 2963.
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| CN101560487B true CN101560487B (en) | 2010-12-08 |
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| CN101985376B (en) * | 2010-06-28 | 2013-05-08 | 北京大学 | Method for removing carbon and nitrogen pollutants in waste water in one step |
| CN102674618B (en) * | 2011-03-10 | 2014-02-12 | 北京科技大学 | High-efficient treatment method for biologically-enhanced coking waste water for biological membrane |
| CN102533623B (en) * | 2012-03-06 | 2013-05-22 | 北京大学 | Achromobacter xylosoxidans with denitrification and dephosphorization function and application of Achromobacter xylosoxidans |
| CN102531202B (en) * | 2012-03-06 | 2013-08-28 | 北京大学 | Comonas testosteroni with denitrification and dephosphorization functions and application thereof |
| CN103421692B (en) * | 2012-05-22 | 2015-02-18 | 上海医药工业研究院 | Compound microorganism live bacteria preparation for enhancing activated sludge, preparation method and applications thereof |
| CN102690762B (en) * | 2012-06-25 | 2013-10-02 | 烟台恒源生物股份有限公司 | Method for producing L-alanine by bioconverting L-aspartic acid by free cell process |
| CN103045509A (en) * | 2012-12-13 | 2013-04-17 | 霍泽仁 | Application of Comamonas testosterone Link |
| CN103146604B (en) * | 2013-02-27 | 2014-01-29 | 中蓝连海设计研究院 | Comamonas testosteroni LH-N5 and heterotrophic nitrification-aerobic denitrification microbial inoculum, and preparation method and application thereof |
| CN103432900B (en) * | 2013-09-07 | 2015-08-12 | 中蓝连海设计研究院 | The purposes of a kind of microbial bacterial agent in foul gas pollution control |
| CN105624063A (en) * | 2016-02-04 | 2016-06-01 | 云南圣清环保科技有限公司 | Efficient mixed flora for treating sludge |
| CN115948288B (en) * | 2022-12-06 | 2023-09-26 | 浙江大学 | Aerobic high-efficiency denitrification compound flora and application thereof |
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