CN102864098A - Denitrification phosphorus removal bacterium H-hrb02 as well as screening method and application thereof - Google Patents
Denitrification phosphorus removal bacterium H-hrb02 as well as screening method and application thereof Download PDFInfo
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Abstract
The invention discloses a denitrification phosphorus removal bacterium as well as a screening method and application efficiency thereof. According to identification, the denitrification phosphorus removal bacterium belongs to pseudomonas, is pseudomonasaeruginosa and is named H-hrb02 and collected in the China General Microbiological Culture Collection Center with the collection number CGMCC NO.5940 on February 27, 2012. An H-hrb02 bacillus is a non-fermented Gram-negative bacterium as well as an obligate aerobe (except that in a nitrate culture medium), grows in the temperature range of 25-42 DEG C, preferably grows at the temperature of 25-30 DEG C, does not grow at the temperature of 4 DEG C particularly, but can grow at the temperature of 42 DEG C. The bacillus is 1.5-5.0 mu m in length and 0.5-1mu m in width, is cue-stick-shaped or linear, and is arranged pairwise or in the form of a short chain; and one end of the bacillus is monotrichous and is free from spores. In a denitrification phosphorus removal strain H-hrb02 provided by the invention, a plurality of organic matters can be taken as carbon sources. After being activated and cultured, the strain (H-hrb02) has excellent denitrification and phosphorus removal performances and can be applied to a sewage biological nitrogen and phosphorus removal process, the phosphorus absorption rate can be up to 92 percent, and the nitrogen removing rate is up to 87.7 percent.
Description
Technical field
The present invention relates to a kind of denitrification phosphorus-collecting bacterium H-hrb02 and effectiveness thereof, the screening method of this bacterium is provided simultaneously.
Background technology
Current, along with national economy develop rapidly with urbanization process deepen continuously, the total emission volumn of urban life and trade effluent is also increasing year by year.And being widely used of agricultural chemicals, chemical fertilizer and synthetic detergent etc., so that the nutritive substance concentration in the water body constantly raises, nitrogen wherein, phosphorus is the one of the main reasons that causes body eutrophication.Conventional biochemical processing process can reduce the BOD in the sewage effectively
5And SS, but work as simultaneous N in the sewage, and when P etc. are nutraceutical, can only remove the 10%-20% of phosphor in sewage, a large amount of phosphorus-containing wastewaters will directly enter environment water.
Traditional biological denitrification phosphorous removal technique exists self insoluble contradiction: polyP bacteria and denitrifying bacteria exist all the time to the competition of carbon source; PolyP bacteria, nitrifier are different with the denitrifying bacteria generation cycle, and the mixing meeting of flora restricts mutually, and system is difficult to the treatment effect that reaches best.Denitrifying Phosphate Accumulating Organisms DPB(Denitrification Phosphorus Removal Bacteria) discovery makes denitrification dephosphorization technique that more vast potential for future development arranged.In case in the middle of sewage treatment process, introduce the PAOs that this class has denitrification function, will produce very important meaning to the technical renovation of biological phosphate-eliminating.At present, characteristic research at the amphimicrobian Denitrifying Phosphate Accumulating Organisms is under the prerequisite on basis, carried out the research of two sludge systems that the poly (hydroxyalkanoate) (PHA) that stores in the organism is separated as the novel process of carbon source and Denitrifying Phosphate Accumulating Organisms and nitrifier, these researchs more and more are subject to scholar's attention.
Up to the present, the denitrification phosphorus-collecting bacterial strain mainly concentrates on
PseudomonasPseudomonas and Bacillus Pseudomonas, bacterial classification is comparatively single, and research focus mostly on aspect degrading genes and mechanism of degradation, real application research is limited.Therefore, filter out having the bacterial strain of efficient denitrification dephosphorization usefulness from nature
PseudomonasaeruginosaStrain H-hrb02, it all is essential carrying out deep research to its physio-biochemical characteristics and denitrogenation dephosphorizing usefulness thereof, has important practical significance and actual application value.
Summary of the invention
One of purpose of the present invention is to provide the pcr amplification process of its DNA in a kind of denitrification phosphorus-collecting bacterium H-hrb02 and the screening method thereof.
Two of purpose of the present invention is to provide the effectiveness of this bacterial strain H-hrb02 in water treatment.
For achieving the above object, the present invention adopts following technical scheme:
One strain denitrification phosphorus-collecting bacterium H-hrb02, it is characterized in that its be Pseudomonas aeruginosa (
Pseudomonas aeruginosa) H-hrb02, belong to Rhodopseudomonas (
Pseudomonas), in Chinese microorganism strain preservation center preservation, preservation date is on 03 27th, 2012, deposit number is CGMCC NO.5940.Denitrification phosphorus-collecting bacterial strain provided by the present invention
PseudomonasaeruginosaStrain H-hrb02 is through being accredited as gram negative bacillus, obligate aerobe (except in nitrate culture-medium), growth temperature range 25-42 ℃, optimum growth temperature is 25-30 ℃, and particularly this bacterium does not grow and can grow at 42 ℃ at 4 ℃, and thalline length is 1.5-5.0
M, width are 0.5-1
M, elongated and different in size, be club shape or wire, paired or short chain shape is arranged, and thalline one end has single flagellum, without gemma.
The screening method of denitrification phosphorus-collecting bacterium H-hrb02, behind the DNA of conventional extraction denitrification phosphorus-collecting bacterium, DNA obtains the base sequence of Denitrifying Phosphate Accumulating Organisms gene through pcr amplification, and the concrete steps of pcr amplification process are:
A. PCR Establishing (25 μ L):
10×PCR Buffer 2.5μL
DNTPs(concentration is 2.5mmol/L) 2 μ L
Primer 1 0.5 μ L
Primer 2 0.5 μ L
Dna profiling 0.5 μ L
RTaq archaeal dna polymerase (5U/ μ L) 0.5 μ L
Add aseptic deionized water to 25 μ L
B. PCR program setting:
95 ℃ of 3min of denaturation
95 ℃ of denaturation 5min, 94 ℃ of sex change 1min, 58 ℃ of renaturation 30s, 72 ℃ are extended 3min, totally 30 circulations, last 72 ℃ are extended 10min;
C. primer sequence:
Primer 15 '-CGCCAGGGTTTTCCCAGTCACGAC-3 '
Primer 25 '-AGCGGATAACAATTTCACACAGGA-3 '.
The purposes of denitrification phosphorus-collecting bacterial strain provided by the present invention is the denitrogenation dephosphorizing efficiency application in the Nitrogen-and Phosphorus-containing wastewater treatment.
Treatment process is: picking H-hrb02 bacterium colony, enrichment are after 24 hours, and the ratio of inoculating 100ml bacterium liquid with every 1L water sample is seeded in scarce phosphorus substratum and the rich phosphorus substratum, and incubation time is 80 hours.
Described scarce phosphorus substratum is formulated as follows (PO
4 3--P is 4mg/L): CH
3COONa 3.23g/L; Na
2HPO
42H
2O 23mg/L; NH
4Cl 152.8mg/L; MgSO
47H
2O 81.12mg/L; K
2SO
417.83mg/L; CaCl
22H
2O 11mg/L; HEPES buffer 7g/L; Trace element 2ml/L; Agar 15g/L; H
2O 1000ml; PH 7.2.121 ℃ of high pressure steam sterilization 20min.
Described rich phosphorus substratum is formulated as follows (PO
4 3--P is 8mg/L): CH
3COONa 3.23g/L; KH
3PO
425mg/L; NH
4Cl 305.52mg/L; MgSO
47H
2O 91.26mg/L; CaCl
22H
2O 25.68mg/L; PIPES buffer 8.5g/L; Trace element 2ml/L; Agar 15g/L; H
2O 1000ml; PH 7.2.121 ℃ of high pressure steam sterilization 20min.
Being formulated as follows of described trace element: FeCl
30.90g/L, H
3BO
30.15g/L, CuSO
45H
2O 0.03g/L, KI 0.18g/L, MnCl
24H
2O 0.06g/L, ZnSO
47H
2O 0.12g/L, CoCl
20.15g/L, Na
2MoO
42H
2O 0.06g/L, EDTA 10.00g/L.
The gene that Pseudomonas aeruginosa provided by the present invention belongs to the H-hrb02 bacterium has outstanding denitrification and poly-phosphorus performance.It can have denitrification and phosphorus removal functional simultaneously, and under proper condition, inhaling the phosphorus rate can be up to 92%, and nitrogen removal efficiency is up to 87.7%, and these characteristics are significant in water treatment field.
Description of drawings
Fig. 1 is the bacterial strain transmission electron microscope observing figure of (Pseudomonas aeruginosa belongs to the H-hrb02 bacterium).
Fig. 2 is the individual atomic power photo of bacterial strain (Pseudomonas aeruginosa belongs to the H-hrb02 bacterium).
Fig. 3 is the bacterial strain growth curve of (Pseudomonas aeruginosa belongs to the H-hrb02 bacterium).
Fig. 4 is that bacterial strain (Pseudomonas aeruginosa belongs to the H-hrb02 bacterium) is inhaled phosphorus usefulness.
Embodiment
Embodiment one: the present embodiment Pseudomonas aeruginosa belong to the H-hrb02 bacterium (
PseudomonasaeruginosaStrain H-hrb02) be to record by following steps:
1. Pseudomonas aeruginosa belongs to separation, purifying and the screening of H-hrb02 bacterium:
A. get the active sludge at sbr reactor device cycle anoxic section end when stable as separation mud
B. adopting dilution to mix flat band method separates.Denitrifying bacteria uses the denitrification substratum to separate, and polyP bacteria uses rich phosphorus substratum to separate.
Concrete separating step is: get the 10mL active sludge to the triangular flask that the 90mL sterilized water is housed from sbr reactor device anoxic section end, add sterile glass beads.Triangular flask is put into shaking table fully vibrate, make bacterium be unicellular state and be scattered in the water.Then, get the access of 1mL suspension the sludge bacteria suspension in triangular flask and the 9mL sterilized water is housed in vitro, obtain the bacteria suspension of 10-1 gradient, continue this step, obtain successively 10-2,10-3,10-4 ... the bacteria suspension of 10-7 gradient is respectively got the 1mL water sample and is put into the culture dish kind, treat to pour in the culture dish when substratum coagulates but soon, mixing after the inversion culture dish is cultivated 2～3 days after the culture medium solidifying in 30 ℃ of constant incubators, is selected the culture dish of a suitable gradient.Then choose respectively clearly colonies typical of different shape, bacterium colony, the single bacterium colony of mark and picking carries out three rides at plate culture medium to be separated, repeat 3～4 times to the consistent single bacterium colony of colony characteristics, the last single bacterium colony of picking, be transferred on the ready test tube slant with for subsequent use, all operations all carries out under aseptic condition.
C. the bacterial strain of oneself being separated is inhaled the phosphorus test in rich phosphorus substratum, assist simultaneously and carry out the test of nitrate reduction aerogenesis and Babes-Ernst bodies dyeing (methylene blue staining method) and PHB particle dyeing (sudan black staining) check.Positive and the aerogenesis for nitrate reductase contains again polyphosphate particle or PHB particle in the thalline, the bacterium of the excessive suction phosphorus of energy is the required Denitrifying Phosphate Accumulating Organisms kind of test under aerobic or anoxia condition.
Concrete test is as follows: aerobic suction phosphorus test: each bacterial strain that oneself is separated at first carries out enrichment culture (seed liquor), be about to bacterial strain to be measured 30 ℃ of anaerobism (inflated with nitrogen) in scarce phosphorus substratum and cultivate 24h, shaking speed is 140rpm(revolutions per minute), to exhaust endobacillary PHB particle as far as possible.The bacterium liquid centrifugal (8,000rpm, 8min) that enrichment culture is good, the sterile distilled water washing, again centrifugal (8,000rpm, 8min) are got the bacterium precipitation and are put in the rich phosphorus substratum, whole process all needs then to carry out aerobic suction phosphorus (140rpm, 30 ℃) test under aseptic condition.Regularly get nutrient solution during this time, behind 0.22 μ m filtering with microporous membrane, detect PO in the filtrate
4 3-The variation of-P concentration.Nitrate reduction aerogenesis test: get each slant strains, be seeded in nitrate reduction substratum (extractum carnis, 3g/L with the Du Shi tubule; Peptone, 5g/L; KNO
3, 1g/L; PH, 7.4; 121 ℃ of high pressure steam sterilization 20min) in, each bacterial strain is done 2 parallel tests, stays in addition 2 pipes not inoculate simultaneously and does contrast.30 ℃ of constant temperature culture, respectively detected result after 1 day, 3 days, 5 days: whether check in the Du Shi tubule to show that if any bubble nitrogen gas generation is arranged by aerogenesis; In the colorimetric porcelain dish, add a small amount of nutrient solution, splash into 1～2 griess reagent A liquid (Sulphanilic Acid, 0.5g; 10% left and right sides dilute acetic acid is 150ml) with B liquid (α-aniline, 0.1g; 10% left and right sides dilute acetic acid, 150ml; H2O, 20ml).Same adding A liquid and B liquid are each 1～2 in control tube.If solution becomes redness, orange, or brown etc., expression has nitrite to exist, and is the nitrate reduction positive; Such as redfree 1～2 pentanoic reagent (the 0.5g pentanoic is dissolved in the dense H2SO4 of 100ml, and adds the 20ml distilled water diluting, is stored in the brown bottle) appears then can adding, as not being blue reaction, then still positive reaction.If be blue reaction, then negative reaction.
2. the physiological and biochemical analysis of bacterial strain (Pseudomonas aeruginosa H-hrb02): the Physiology and biochemistry of bacterial strain (Pseudomonas aeruginosa H-hrb02) is identified with reference to " the 8th edition operation of the outstanding Bacteria Identification handbook of uncle, qualification result is that to belong to the H-hrb02 bacterium be non-fermentation Gram-negative bacteria to Pseudomonas aeruginosa, obligate aerobe (except in nitrate culture-medium), growth temperature range 25-42 ℃, optimum growth temperature is 25-30 ℃, and particularly this bacterium does not grow and can grow at 42 ℃ at 4 ℃.Thalline length is 1.5-5.0
M, width are 0.5-1
M is club shape or wire, and paired or short chain shape is arranged, and thalline one end has single flagellum, without gemma.This bacterial strain is without the need for the machine growth factor.The nutrition variation; The growth of single bacterial strain can utilize 76-82 kinds or more different organic compound.Bacterial strain is carried out gramstaining, oxydase, catalase, indoles is produced in the glucose oxidase fermentation, and V.P. measures, and M.R. measures, gelatine liquefication, Starch Hydrolysis, the Citrate trianion utilization, the test of the major physiological biochemical indicators such as nitrate reduction, the result is as shown in table 2.
In the present embodiment step 1, the denitrification substratum is formulated as follows: Trisodium Citrate 5.0g/L; K
2HPO
41.0g/L; KH
2PO
41.0g/L; MgSO
47H
2O 0.2g/L; KNO
32.0g/L, agar 15g/L.pH 7.2。121 ℃ of high pressure steam sterilization 20min.
Embodiment two: the 16S rDNA order-checking of gene, the pcr amplification concrete steps of 16S rDNA gene are as follows:
A. PCR Establishing (25 μ L):
10×PCR Buffer 2.5μL
DNTPs(concentration is 2.5mmol/L) 2 μ L
Primer 1 0.5 μ L
Primer 2 0.5 μ L
Dna profiling 0.5 μ L
RTaq archaeal dna polymerase (5U/ μ L) 0.5 μ L
Add aseptic deionized water to 25 μ L
B. PCR program setting:
95 ℃ of 3min of denaturation
95 ℃ of denaturation 5min, 94 ℃ of sex change 1min, 58 ℃ of renaturation 30s, 72 ℃ are extended 3min, totally 30 circulations, last 72 ℃ are extended 10min;
C. primer sequence:
Primer 15 '-CGCCAGGGTTTTCCCAGTCACGAC-3 '
Primer 25 '-AGCGGATAACAATTTCACACAGGA-3 '
Pseudomonas aeruginosa H-hrb02 gene order is shown in SEQ IDNo.1.
Embodiment three: present embodiment Pseudomonas aeruginosa H-hrb02 can take gas chromatography as carbon source, can be used for the bio-denitrifying sewage dephosphorization.Pseudomonas aeruginosa H-hrb02 biological carbon and phosphorous removal ability is detected:
1. Pseudomonas aeruginosa H-hrb02 growth curve is measured.According to the standard method of measuring growth curve of bacteria the growth curve of bacterial strain (Pseudomonas aeruginosa H-hrb02) is measured, got nutrient solution every 2h, adopt photoelectric turbidimetry, measure the OD of bacterium liquid at 600nm wavelength place
600(Optical Density) then through 0.22 μ m filtering with microporous membrane, detects filtrate PO
4 3--P, the indexs such as pH value.Obtain Pseudomonas aeruginosa H-hrb02 growth curve as shown in Figure 3.As can be seen from Figure 3, the growth curve of this bacterial strain (Pseudomonas aeruginosa H-hrb02) is the typical case relatively, latent period is shorter, for about 2h, this may be because the activity of bacterial strain (Pseudomonas aeruginosa H-hrb02) is stronger, and the culture condition before and after the inoculation is similar, and can adapt to new environment within a short period of time after the access.The logarithmic phase of bacterial strain is about 14～16h, and 18h begins later on to enter stationary phase and decline phase.Just begin to inhale phosphorus after also can learning in bacterial strain (Pseudomonas aeruginosa H-hrb02) the access nutrient solution from figure, but mainly occur in logarithmic phase, in this period, the microorganism growth metabolism is the most vigorous, inhales the phosphorus amount and increases gradually along with the growth of logarithmic phase bacterial strain.Inhale the phosphorus rate and after bacterial strain enters stationary phase, reach maximum, then occurred releasing the phosphorus phenomenon, but it is very little to release the phosphorus amount.
2. Pseudomonas aeruginosa H-hrb02 denitrogenation dephosphorizing potency assay
A. Pseudomonas aeruginosa H-hrb02 denitrification efficiency is measured:
Bacterial strain (Pseudomonas aeruginosa H-hrb02) is activated enrichment culture, bacteria suspension centrifugal (8 according to embodiment one method, 000rpm, 10min), abandon supernatant liquor, the bacterium precipitation is forwarded in the rich phosphorus substratum that does not contain sodium acetate, cultivates 48h under good oxygen condition.As can be seen from Table 1, bacterial strain H-hrb02 has good aerobic denitrification effect.In general the conventional denitrification theory is thought, because dissolved oxygen meeting and NO
3-The competition electron acceptor(EA), the existence meeting of dissolved oxygen (DO) is played restraining effect to denitrification process, carries out thereby suppress denitrifying.But the proposition of aerobic denitrification theory has changed traditional concept, and its theory is thought owing to having the pericentral siphon nitrate reductase in the thalline, so bacterial strain can utilize nitric nitrogen and oxygen to carry out co-respiration as electron acceptor(EA) simultaneously.To sum up, bacterial strain H-hrb02 is efficient Denitrifying Phosphate Accumulating Organisms.
B. Pseudomonas aeruginosa H-hrb02 inhales the phosphorus potency assay:
Bacterial strain (Pseudomonas aeruginosa H-hrb02) is activated according to embodiment one method, enrichment culture, bacteria suspension centrifugal (8,000rpm, 10min), abandon supernatant liquor, it is 2.5 that the bacterium precipitation is forwarded to rich phosphorus substratum dissolved oxygen, and for guaranteeing ambient stable in the substratum, the pH value of avoiding bacterial strain to cause in inhaling the phosphorus process fluctuates in a big way, add HEPES(hydrogen ion buffer reagent in the substratum), it act as the pH scope of long period in controling environment.As seen from Figure 4, bacterial strain H-hrb02 inhales the phosphorus rate and reaches 85% when 12h.Illustrate that phosphor-removing effect is good in the bacterial strain H-hrb02 short period of time.But by finding out among Fig. 4 that the prolongation dephosphorizing rate along with incubation time descends gradually, the phosphorus phenomenon has appearred releasing in bacterial strain H-hrb02.Analyze its reason and may be the prolongation along with the reaction times, available NO
3--N is not enough, causes bacterial strain not have enough electron acceptor(EA)s so that dephosphorizing rate descends.Another reason may be that the carbon source deficiency can not provide nutritive substance and energy for microbial growth and breeding in the substratum.And enough organic substances are the prerequisites of the synthetic PHB of polyP bacteria, just directly have influence on the growth and reproduction of Denitrifying Phosphate Accumulating Organisms in the time can not synthesizing PHB, thereby have suppressed dephosphorization efficiency by using.
The dna sequence dna table
Pseudomonas aeruginosa H-hrb02 16S rDNA SEQ. ID. No.1:
1 gattgcggac gggtgagtaa tgcctaggaa tctgcctggt agtgggggat aacgtccgga
61 aacgggcgct aataccgcat acgtcctgag ggagaaagtg ggggatcttc ggacctcacg
121 ctatcagatg agcctaggtc ggattagcta gttggtgggg taaaggccta ccaaggcgac
181 gatccgtaac tggtctgaga ggatgatcag tcacactgga actgagacac ggtccagact
241 cctacgggag gcagcagtgg ggaatattgg acaatgggcg aaagcctgat ccagccatgc
301 cgcgtgtgtg aagaaggtct tcggatcgta aagcacttta agttgggagg aagggcagta
361 agttaatacc ttgctgtttt gacgttacca acagaataag caccggctaa cttcgtgcca
421 gcagccgcgg taataat
1 gattgcggac gggtgagtaa tgcctaggaa tctgcctggt agtgggggat aacgtccgga
61 aacgggcgct aataccgcat acgtcctgag ggagaaagtg ggggatcttc ggacctcacg
121 ctatcagatg agcctaggtc ggattagcta gttggtgggg taaaggccta ccaaggcgac
181 gatccgtaac tggtctgaga ggatgatcag tcacactgga actgagacac ggtccagact
241 cctacgggag gcagcagtgg ggaatattgg acaatgggcg aaagcctgat ccagccatgc
301 cgcgtgtgtg aagaaggtct tcggatcgta aagcacttta agttgggagg aagggcagta
361 agttaatacc ttgctgtttt gacgttacca acagaataag caccggctaa cttcgtgcca
421 gcagccgcgg taataat
Claims (5)
1. a strain denitrification phosphorus-collecting bacterium H-hrb02, it is characterized in that its be Pseudomonas aeruginosa (
Pseudomonas aeruginosa) H-hrb02, belong to Rhodopseudomonas (
Pseudomonas), in Chinese microorganism strain preservation center preservation, preservation date is on 03 27th, 2012, deposit number is CGMCC NO.5940.
2. denitrification phosphorus-collecting bacterium H-hrb02 according to claim 1 is characterized in that bacterial strain
PseudomonasaeruginosaStrain H-hrb02 is through being accredited as gram negative bacillus, obligate aerobe (except in nitrate culture-medium), growth temperature range 25-42 ℃, optimum growth temperature is 25-30 ℃, particularly this bacterium does not grow and can grow at 42 ℃ at 4 ℃, and thalline length is 1.5-5.0
M, width are 0.5-1
M is club shape or wire, and paired or short chain shape is arranged, and thalline one end has single flagellum, without gemma.
3. the screening method of denitrification phosphorus-collecting bacterium H-hrb02 according to claim 1 and 2, behind the DNA of conventional extraction denitrification phosphorus-collecting bacterium, DNA obtains the base sequence of Denitrifying Phosphate Accumulating Organisms gene through pcr amplification, it is characterized in that the concrete steps of pcr amplification process are:
A. PCR Establishing (25 μ L):
10×PCR Buffer 2.5μL
DNTPs(concentration is 2.5mmol/L) 2 μ L
Primer 1 0.5 μ L
Primer 2 0.5 μ L
Dna profiling 0.5 μ L
RTaq archaeal dna polymerase (5U/ μ L) 0.5 μ L
Add aseptic deionized water to 25 μ L
B. PCR program setting:
95 ℃ of 3min of denaturation
95 ℃ of denaturation 5min, 94 ℃ of sex change 1min, 58 ℃ of renaturation 30s, 72 ℃ are extended 3min, totally 30 circulations, last 72 ℃ are extended 10min;
C. primer sequence:
Primer 15 '-CGCCAGGGTTTTCCCAGTCACGAC-3 '
Primer 25 '-AGCGGATAACAATTTCACACAGGA-3 '.
4. the denitrogenation dephosphorizing efficiency application of denitrification phosphorus-collecting bacterium H-hrb02 according to claim 1 and 2 in the Nitrogen-and Phosphorus-containing wastewater treatment.
5. the denitrogenation dephosphorizing efficiency application of denitrification phosphorus-collecting bacterium H-hrb02 according to claim 4, it is characterized in that treatment process is: picking H-hrb02 bacterium colony, after the enrichment 24 hours, ratio with every 1L water sample inoculation 100ml bacterium liquid is seeded in scarce phosphorus substratum and the rich phosphorus substratum, and incubation time is 80 hours;
Described scarce phosphorus substratum be formulated as follows (PO
4 3--P is 4mg/L): CH
3COONa 3.23g/L, Na
2HPO
42H
2O 23mg/L, NH
4Cl 152.8mg/L, MgSO
47H
2O 81.12mg/L, K
2SO
417.83mg/L, CaCl
22H
2O 11mg/L, HEPES buffer 7g/L, micro-2ml/L, agar 15g/L, H
2O 1000ml, 7.2,121 ℃ of high pressure steam sterilization 20min of pH;
Described rich phosphorus substratum be formulated as follows (PO
4 3--P is 8mg/L): CH
3COONa 3.23g/L, KH
3PO
425mg/L, NH
4Cl 305.52mg/L, MgSO
47H
2O 91.26mg/L, CaCl
22H
2O 25.68mg/L, PIPES buffer 8.5g/L, micro-2ml/L, agar 15g/L, H
2O 1000ml, 7.2,121 ℃ of high pressure steam sterilization 20min of pH;
Being formulated as follows of described trace element: FeCl
30.90g/L, H
3BO
30.15g/L, CuSO
45H
2O 0.03g/L, KI 0.18g/L, MnCl
24H
2O 0.06g/L, ZnSO
47H
2O 0.12g/L, CoCl
20.15g/L, Na
2MoO
42H
2O 0.06g/L, EDTA 10.00g/L.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103114062A (en) * | 2013-01-29 | 2013-05-22 | 山东大学 | Denitrifying phosphate-accumulating organism with nitrogen and phosphorus removal functions and applications thereof |
| CN104212750A (en) * | 2014-09-17 | 2014-12-17 | 常州大学 | Phosphorus-accumulating bacterium culture medium and preparation method thereof |
| CN106244502A (en) * | 2016-09-27 | 2016-12-21 | 湖北大学 | One strain efficient dephosphorization and the pseudomonas of degraded lecithin |
| CN109486699A (en) * | 2018-08-21 | 2019-03-19 | 四川大学 | One plant of salt tolerant aerobic denitrifying bacteria and application |
| CN114480159A (en) * | 2021-11-23 | 2022-05-13 | 北京工业大学 | Synchronous heterotrophic nitrification aerobic denitrification phosphorus removal bacterium and application thereof |
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| CN103114062A (en) * | 2013-01-29 | 2013-05-22 | 山东大学 | Denitrifying phosphate-accumulating organism with nitrogen and phosphorus removal functions and applications thereof |
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| CN104212750A (en) * | 2014-09-17 | 2014-12-17 | 常州大学 | Phosphorus-accumulating bacterium culture medium and preparation method thereof |
| CN106244502A (en) * | 2016-09-27 | 2016-12-21 | 湖北大学 | One strain efficient dephosphorization and the pseudomonas of degraded lecithin |
| CN106244502B (en) * | 2016-09-27 | 2019-06-18 | 湖北大学 | The pseudomonad of one plant of efficient dephosphorization and degradation lecithin |
| CN109486699A (en) * | 2018-08-21 | 2019-03-19 | 四川大学 | One plant of salt tolerant aerobic denitrifying bacteria and application |
| CN114480159A (en) * | 2021-11-23 | 2022-05-13 | 北京工业大学 | Synchronous heterotrophic nitrification aerobic denitrification phosphorus removal bacterium and application thereof |
| CN114480159B (en) * | 2021-11-23 | 2024-03-29 | 北京工业大学 | Synchronous heterotrophic nitrification aerobic denitrification dephosphorization bacterium and application thereof |
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