CN103805546A - Acinetobacter johnsonii AJ-3 strain and application thereof - Google Patents
Acinetobacter johnsonii AJ-3 strain and application thereof Download PDFInfo
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- CN103805546A CN103805546A CN201410074492.9A CN201410074492A CN103805546A CN 103805546 A CN103805546 A CN 103805546A CN 201410074492 A CN201410074492 A CN 201410074492A CN 103805546 A CN103805546 A CN 103805546A
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- 241000122229 Acinetobacter johnsonii Species 0.000 title claims abstract description 30
- 239000010865 sewage Substances 0.000 claims abstract description 19
- 230000001580 bacterial effect Effects 0.000 claims description 44
- 238000004321 preservation Methods 0.000 claims description 7
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 abstract description 32
- 229910052698 phosphorus Inorganic materials 0.000 abstract description 32
- 239000011574 phosphorus Substances 0.000 abstract description 32
- 238000000034 method Methods 0.000 abstract description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 13
- 230000012010 growth Effects 0.000 abstract description 12
- 230000008569 process Effects 0.000 abstract description 4
- 230000007613 environmental effect Effects 0.000 abstract description 3
- 238000000926 separation method Methods 0.000 abstract description 3
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- 230000000813 microbial effect Effects 0.000 abstract description 2
- 244000005700 microbiome Species 0.000 abstract description 2
- 229910052760 oxygen Inorganic materials 0.000 abstract description 2
- 239000001301 oxygen Substances 0.000 abstract description 2
- 239000001963 growth medium Substances 0.000 abstract 1
- 238000009630 liquid culture Methods 0.000 abstract 1
- 239000007788 liquid Substances 0.000 description 17
- 241000894006 Bacteria Species 0.000 description 15
- 239000001888 Peptone Substances 0.000 description 15
- 235000015278 beef Nutrition 0.000 description 13
- 239000007787 solid Substances 0.000 description 12
- 235000015097 nutrients Nutrition 0.000 description 11
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- 229920000388 Polyphosphate Polymers 0.000 description 8
- 229920000037 Polyproline Polymers 0.000 description 8
- 239000001205 polyphosphate Substances 0.000 description 8
- 235000011176 polyphosphates Nutrition 0.000 description 8
- 108020004465 16S ribosomal RNA Proteins 0.000 description 6
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- 238000012360 testing method Methods 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 5
- 239000010802 sludge Substances 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 3
- 229930064664 L-arginine Natural products 0.000 description 3
- 235000014852 L-arginine Nutrition 0.000 description 3
- 229910002651 NO3 Inorganic materials 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- APUPEJJSWDHEBO-UHFFFAOYSA-P ammonium molybdate Chemical compound [NH4+].[NH4+].[O-][Mo]([O-])(=O)=O APUPEJJSWDHEBO-UHFFFAOYSA-P 0.000 description 3
- 235000018660 ammonium molybdate Nutrition 0.000 description 3
- 239000011609 ammonium molybdate Substances 0.000 description 3
- 229940010552 ammonium molybdate Drugs 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000009294 enhanced biological phosphorus removal Methods 0.000 description 3
- 238000012851 eutrophication Methods 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 229920000159 gelatin Polymers 0.000 description 3
- 235000019322 gelatine Nutrition 0.000 description 3
- BHEPBYXIRTUNPN-UHFFFAOYSA-N hydridophosphorus(.) (triplet) Chemical compound [PH] BHEPBYXIRTUNPN-UHFFFAOYSA-N 0.000 description 3
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 3
- 150000002475 indoles Chemical class 0.000 description 3
- 239000002054 inoculum Substances 0.000 description 3
- CEQFOVLGLXCDCX-WUKNDPDISA-N methyl red Chemical compound C1=CC(N(C)C)=CC=C1\N=N\C1=CC=CC=C1C(O)=O CEQFOVLGLXCDCX-WUKNDPDISA-N 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000002798 spectrophotometry method Methods 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 241000589291 Acinetobacter Species 0.000 description 2
- 201000004384 Alopecia Diseases 0.000 description 2
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 2
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- 241000726221 Gemma Species 0.000 description 2
- ODKSFYDXXFIFQN-BYPYZUCNSA-N L-arginine Chemical compound OC(=O)[C@@H](N)CCCN=C(N)N ODKSFYDXXFIFQN-BYPYZUCNSA-N 0.000 description 2
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 239000002351 wastewater Substances 0.000 description 2
- 239000001828 Gelatine Substances 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 108010077895 Sarcosine Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- YCUVUDODLRLVIC-UHFFFAOYSA-N Sudan black B Chemical compound C1=CC(=C23)NC(C)(C)NC2=CC=CC3=C1N=NC(C1=CC=CC=C11)=CC=C1N=NC1=CC=CC=C1 YCUVUDODLRLVIC-UHFFFAOYSA-N 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- QGAVSDVURUSLQK-UHFFFAOYSA-N ammonium heptamolybdate Chemical compound N.N.N.N.N.N.O.O.O.O.O.O.O.O.O.O.O.O.O.O.O.O.O.O.O.O.O.O.O.O.[Mo].[Mo].[Mo].[Mo].[Mo].[Mo].[Mo] QGAVSDVURUSLQK-UHFFFAOYSA-N 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
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- OKKJLVBELUTLKV-UHFFFAOYSA-N methanol Natural products OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 1
- CXKWCBBOMKCUKX-UHFFFAOYSA-M methylene blue Chemical compound [Cl-].C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 CXKWCBBOMKCUKX-UHFFFAOYSA-M 0.000 description 1
- 229960000907 methylthioninium chloride Drugs 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
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- 229920013639 polyalphaolefin Polymers 0.000 description 1
- 229920000137 polyphosphoric acid Polymers 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- FSYKKLYZXJSNPZ-UHFFFAOYSA-N sarcosine Chemical compound C[NH2+]CC([O-])=O FSYKKLYZXJSNPZ-UHFFFAOYSA-N 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the field of environmental microorganisms and in particular relates to separation and culture of an Acinetobacter johnsonii AJ-3 strain and application thereof in phosphorus removal in sewage and biological phosphorus removal in urban sewage treatment plants. The Acinetobacter johnsonii AJ-3 strain has a collection number of CCTCCM2014023, the collection place is China Center for Type Culture Collection, and the collection date is January, 16th 2014. The strain is an Acinetobacter johnsonii AJ-3 strain, is rapid in growth, high in adaptive capacity to environment and low in dissolved oxygen demands and can rapidly grow in a phosphorus-containing synthetic sewage liquid culture medium, the phosphorus is removed, the phosphorus removal efficiency is high and stable and reaches 55-60.1%, and secondary pollution is avoided. Therefore, the AJ-3 strain can be applied to phosphorus-containing sewage treatment, is an important microbial resource in a water organism phosphorus removal process and has wide application prospect.
Description
Technical field
The invention belongs to environmental microorganism field
,be specifically related to a strain Acinetobacter johnsonii (
acinetobacter johnsonii) separation, cultivation and the application in sewage dephosphorization thereof of AJ-3 bacterial strain, be applied to the biological phosphate-eliminating of municipal sewage plant.
Background technology
From 2010 to 2012, China Environmental State Bulletin showed, 62 state control emphasis lakes (reservoir) body eutrophication outstanding problem, and East Sea immediate offshore area water-quality is extreme difference and serious pollution over nearly 3 years, in main contamination index, all comprises phosphorus; And eutrophication will reduce the transparency of water body, produce obnoxious flavour and biotoxin, affect people and animals' health, destroy species diversity in water body, impact tourism and shipping etc.; Therefore, the phosphorus in water body is removed particularly important to preventing body eutrophication.
Current water body dephosphorized technology is mainly divided two classes: chemical dephosphorization and biological phosphate-eliminating.Chemical dephosphorization is mainly chemical precipitation method, have simple to operate, clearance is high, the advantage such as stable; But the chemical sludge amount that this method exists, and dosage is large, processing costs is high, produce is large and be rich in organic matter, and the ultimate disposal of chemical sludge is also a problem.Biological phosphate-eliminating is mainly to utilize the inrichment of polyP bacteria to phosphorus, the mode that remains rich phosphorous sludge by eliminating is removed the phosphorus in water body effectively, the advantages such as biological phosphate-eliminating has that sludge output is few, save energy, working cost are lower, non-secondary pollution are the more phosphorus removing methods of application at present.The conventional sewage dephosphorization method of extensive sewage work is mainly biological dephosphorization, wherein Enhanced Biological Phosphorus Removal method (Enhanced Biological Phosphorus Removal, EBPR) there is sludge yield few, do not use the feature such as chemical substance and economical operation.
PolyP bacteria (Phosphorus Accumulataing Organisms, PAOs) is not the concept in systematic bacteriology, is the general name of the quasi-microorganism to having " excess suction phosphorus " feature.So-called " excess suction phosphorus ", refers to that they can stores unnecessary phosphorus in vivo with the form of polyphosphoric acid salt, prepares against energy and nutrition are provided in poor environment.So the phosphorus content of some polyP bacteria thalline can reach the more than 10% of dry weight.And because can make the phosphorus content in external environment obviously reduce in the process of " excess suction phosphorus ", polyP bacteria just becomes the main executive of biological removal of phosphorus in wastewater, plays a decisive role in biological phosphate-eliminating.
The present invention from natural habitat, be sieved to one select good strains in the field for seed have typical anaerobism characteristic polyP bacteria Acinetobacter johnsonii (
acinetobacter johnsonii) AJ-3 bacterial strain, be intended to abundant polyP bacteria resource, expand polyP bacteria strain library, for trade effluent, biological removal of phosphorus in wastewater provide bacterial classification.
Summary of the invention
The object of this invention is to provide plant height effect polyP bacteria Acinetobacter johnsonii and an application thereof.
Acinetobacter johnsonii of the present invention (
acinetobacter johnsonii) AJ-3 bacterial strain, deposit number is; CCTCC M 2014023, is preservation: Chinese Typical Representative culture collection center, preservation day is: on 01 16th, 2014.
Bacterial strain screening involved in the present invention is from the pedotheque of Zhejiang Normal University of Jinhua, Zhejiang Province city north gate.Adopt gradient dilution method to obtain pure bacterial strain, according to morphological specificity, physiological and biochemical property, 16S rDNA gene order identify AJ-3 bacterial strain be Acinetobacter johnsonii (
acinetobacter johnsonii).
AJ-3 bacterial strain bacterium colony circle, white, neat in edge, protuberance, smooth moistening, translucent (accompanying drawing 1); Thalline is rod-short, Gram-negative, and atrichia, without gemma (accompanying drawing 2).
AJ-3 bacterial strain energy hydrolyzed starch, nitrate reduction and the test of L-arginine decarboxylase are all positive, and can not utilize Citrate trianion, not liquefy gelatin, VP, methyl red, indoles, hydrogen sulfide, all negative (table 1) of glucose fermentation test; Strain growth temperature range is 15 ℃ ~ 40 ℃, and optimum growth temperature is 28 ℃, and suitable growth pH is 7.0 ~ 8.0.
16S rDNA sequencing analysis result shows, its base sequence total length is 1431 bp(accompanying drawings 3), with Acinetobacter johnsonii (
acinetobacter johnsonii) homology be 98% (accompanying drawing 4).
The condition of biological phosphate-eliminating that AJ-3 bacterial strain involved in the present invention can be used for synthetic sewage is as follows: going bail for is stored in the glycerol stock 20 μ L of-20 ℃ and coats on the flat board of beef extract-peptone solid medium, after 28 ℃ of cultivation 24 h, choosing single bacterium colony mixes in 0.3 mL sterilized water, getting 100 μ L coats on the flat board of beef extract-peptone solid medium, getting 2 holes with punch tool (aperture 0.5 cm) receives in 15 mL beef extract-peptone liquid nutrient mediums, be placed in 28 ℃, in 170 r/min shaking tables, cultivate 24 h(OD
600≈ 0.9) activation, make seed liquor; With the synthetic sewage liquid nutrient medium of inoculum size access of 8 %, 28 ℃ of temperature, shaking table speed 170 r/min, cultivate 9 h under the condition that initial pH is 7.2 again.Get 50 mL bacteria suspensions in the centrifuge tube of 50 mL, with centrifugal 5 min of 12000 r/min, to get supernatant liquor and survey phosphorus concentration by the measuring method of total phosphorus, the mensuration of total phosphorus adopts ammonium molybdate spectrophotometry, calculate dephosphorizing rate according to contrast, obtaining the highest dephosphorizing rate is 60.1%(accompanying drawing 6).
The present invention be a strain Acinetobacter johnsonii (
acinetobacter johnsonii) AJ-3 bacterial strain, this strain growth is quick, adaptive capacity to environment is strong, dissolved oxygen is required low, can be in phosphorous synthetic sewage liquid nutrient medium Fast Growth, and phosphorus is removed, dephosphorization efficiency by using is high and stable, reaches 55% ~ 60.1%, non-secondary pollution.Therefore, AJ-3 bacterial strain can be applicable to phosphorous sewage disposal, is Microbial resources important on water body Biological Phosphorus Removal Processes, has a good application prospect.
Accompanying drawing explanation
Fig. 1 is the colony characteristics of AJ-3 bacterial strain.
Fig. 2 is the thalli morphology (× 1000) of AJ-3 bacterial strain.
Fig. 3 is AJ-3 bacterial strain 16S rDNA gene order.
Fig. 4 is the acinetobacter phylogenetic tree of setting up based on 16s rDNA gene order.
Fig. 5 is the growth curve of AJ-3 bacterial strain.
Fig. 6 is the impact of liquid amount on AJ-3 bacterial strain dephosphorizing rate.
biological preservation statement
Acinetobacter johnsonii (
acinetobacter johnsonii) AJ-3 bacterial strain, deposit number is; CCTCC M 2014023, is preservation: Chinese Typical Representative culture collection center, preservation day is: on 01 16th, 2014.
Embodiment
separation screening and the evaluation of the efficient polyP bacteria Acinetobacter johnsonii AJ-3 bacterial strain of embodiment 1
(1) substratum
Beef extract-peptone solid medium: extractum carnis 3 g/L, peptone 10 g/L, NaCl 5 g/L, agar 20 g/L, pH 7.0-7.2 (liquid nutrient medium does not add agar);
Limit phosphorus (rich phosphorus) solid medium: get 30 mL deionized water dissolving 8.372 g 3-morpholine propanesulfonic acid, 0.717g N-tri-(methylol) methylglycine, with the KOH adjusting pH to 7.4 of 10 mol/L, add in the following order solution: 0.01 mL 1.830% FeSO
4solution (now joining), 5 mL 1.9 mol/L NH
4cl, 1 mL 0.276 mol/L K
2sO
4, 0.025 mL 0.02 mol/L CaCl
22H
2o, 0.42 mL 1.25 mol/L MgCl
26 H
2o, 20 mL 2.5 mol/L NaCl, 0.02 mL trace element mixed solution (Ammonium Heptamolybdate 3 × 10
-6mol/L, H
3bO
34 × 10
-4mol/L, CoCl
23 × 10
-5mol/L, CuSO
410
-5mol/L, MnCl
28 × 10
-5mol/L, ZnSO
410
-5mol/L), 10 mL 1% glucose, 0.4 mL 1.686% VITMAIN B1,250 μ L 6.928% K
2hPO
4(rich phosphorus adds 5 mL)
,50 mg para-totuidine indigo plants, add water and are settled to 500 mL, after filtration sterilization, mix and make solid medium be dissolved with 20 g agar and the not solidified 500 mL solution of sterilising treatment.
(2) method
Adopt gradient dilution partition method.Soil sampling 1.0 g, add 9 mL sterilized water dilutions, carry out 10
-1~ 10
-6gradient dilution, gets 10
-5diluent 100 μ L coat on beef extract-peptone solid medium flat board, cultivate 24 h for 28 ℃.The single bacterium colony of picking bacterium, separates, purifying; Be further purified bacterial strain and adopt method of scoring, the pure bacterial strain of gained is put in-20 ℃ of refrigerators and is preserved, for subsequent use.
Bacterial strain is screened with Lv Shi methylene blue staining, sudan black staining, blue hickie method and ammonium molybdate spectrophotometry, after primary dcreening operation and multiple sieve, obtain the bacterial isolates (accompanying drawing 2) that a strain is numbered AJ-3 efficient dephosphorization.
AJ-3 bacterial strain is carried out to morphology, Physiology and biochemistry and 16S r DNA sequence dna to be identified.
(3) result
Strain morphology feature: bacterium colony circle, white, neat in edge, protuberance, smooth moistening, translucent (accompanying drawing 1); Thalline is rod-short, and gramstaining is negative, and atrichia, without gemma (accompanying drawing 2).
Bacterial strain physiological and biochemical property: AJ-3 bacterial strain energy hydrolyzed starch, nitrate reduction and the test of L-arginine decarboxylase are all positive, can not utilize Citrate trianion, not liquefy gelatin, VP, methyl red, indoles, hydrogen sulfide, all negative (table 1) of glucose fermentation test; Strain growth temperature range is 15 ℃ ~ 40 ℃, and optimum growth temperature is 28 ℃, and suitable growth pH is 7.0 ~ 8.0.
16S rDNA gene order (accompanying drawing 3): 16S rDNA gene sequencing analytical results shows, its base sequence total length is 1431 bp, use online Blast software to carry out homology analysis, choose the strain construction phylogenetic tree (accompanying drawing 4) of 21 strain acinetobacters.Shown by Fig. 4, AJ-3 bacterial strain and Acinetobacter johnsonii (
acinetobacter johnsonii) on same of evolutionary tree, two bacterial strain homologys are up to 98%.In conjunction with strain morphology feature, physiological and biochemical property, identify this bacterial strain be Acinetobacter johnsonii (
acinetobacter johnsonii).
The physiological and biochemical test result of table 1 AJ-3 bacterial strain
| Test subject | Result | Test subject | Result |
| Starch Hydrolysis | + | VP | - |
| Nitrate reduction | + | Methyl red | - |
| L-arginine decarboxylase | + | Indoles | - |
| Citrate trianion | - | Hydrogen sulfide | - |
| Gelatine liquefication | - | Glucose fermentation | - |
Note: "+" is positive, "-" is negative.
the growth curve of embodiment 2 Acinetobacter johnsonii AJ-3 bacterial strains is measured
(1) bacterial strain:aJ-3 bacterial strain.
(2) substratum
Beef extract-peptone solid medium: formula is with embodiment 1
Synthetic sewage liquid nutrient medium: sodium acetate 0.925 g/L, peptone 0.1 g/L, yeast extract paste 0.01 g/L, NaCl 0.05 g/L, KH
2pO
40.0655 g/L, MgCl
26H
2o 0.1412 g/L, CaCl
20.025 g/L, pH7.0-7.2.
(3) method
Go bail for and be stored in the glycerol stock 20 μ L of-20 ℃ and coat on the flat board of beef extract-peptone solid medium, after 28 ℃ of cultivation 24 h, choosing single bacterium colony mixes in 0.3 mL sterilized water, getting 100 μ L coats on the flat board of beef extract-peptone solid medium, getting 2 holes with punch tool (aperture 0.5 cm) receives in 15 mL beef extract-peptone liquid nutrient mediums, be placed in 28 ℃, in 170 r/min shaking tables, cultivate 24 h(OD
600≈ 0.9) activation, make seed liquor; Be equipped with in 250 mL Erlenmeyer flasks of the synthetic sewage liquid nutrient medium of 100 mL with 8% inoculum size access again, be placed in 28 ℃, in 170 r/min shaking tables, cultivate, measure OD every 1 h
600value.
(4) result
After AJ-3 bacterial strain is activated, in synthetic sewage substratum, growth is fast, adaptable; In culturing process, do not occur obvious lag phase, 1 h ~ 9 h is logarithmic phase vegetative period, and 9 h ~ 24 h are always in stable growth phase (accompanying drawing 5).
the impact of embodiment 3 liquid amounts on Acinetobacter johnsonii AJ-3 bacterial strain biological phosphor-removing effect.
(1)
bacterial strain:aJ-3 bacterial strain.
(2) substratum
Beef extract-peptone solid medium: formula is with embodiment 1;
Synthetic sewage liquid nutrient medium: formula is with embodiment 2.
(3) method
Go bail for and be stored in the glycerol stock 20 μ L of-20 ℃ and coat on the flat board of beef extract-peptone solid medium, after 28 ℃ of cultivation 24 h, choosing single bacterium colony mixes in 0.3 mL sterilized water, getting 100 μ L coats on the flat board of beef extract-peptone solid medium, getting 2 holes with punch tool (aperture 0.5 cm) receives in 15 mL beef extract-peptone liquid nutrient mediums, be placed in 28 ℃, in 170 r/min shaking tables, cultivate 24 h(OD
600≈ 0.9) activation, make seed liquor; Be equipped with in synthetic sewage liquid nutrient medium (liquid amount is respectively 40 mL, 80 mL, 100 mL, 120 mL, 150 mL, 200 mL) the 250 mL triangular flasks of different amounts with 8% inoculum size access again, 28 ℃ of temperature, shaking table speed 170 r/min, cultivate 9 h under the condition that initial pH is 7.2.Get 25 mL bacteria suspensions in the centrifuge tube of 50 mL, with centrifugal 5 min of 12 000 r/min, get supernatant liquor and survey phosphorus concentration by the measuring method of total phosphorus, the mensuration of total phosphorus adopts ammonium molybdate spectrophotometry, calculates dephosphorizing rate according to contrast.
(4) result
Different liquid amounts have a certain impact to AJ-3 bacterial strain phosphor-removing effect, when liquid amount is to fill 120 mL in 250 mL triangular flasks, and 28 ℃ of temperature, shaking table speed 170 r/min, synthetic sewage liquid nutrient medium (pH is 7.2) is lower cultivates 9 h, and dephosphorizing rate can reach 60.1%; The suitable liquid amount of AJ-3 bacterial strain is 80 mL ~ 150 mL (accompanying drawings 6).
Claims (2)
- Acinetobacter johnsonii ( acinetobacter johnsonii) AJ-3 bacterial strain, it is characterized in that: the deposit number of this bacterial strain is; CCTCC M 2014023, is preservation: Chinese Typical Representative culture collection center, preservation day is: on 01 16th, 2014.
- 2. the purposes of Acinetobacter johnsonii AJ-3 bacterial strain according to claim 1, is characterized in that: this bacterial strain is for the biological phosphate-eliminating of sewage.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105586294A (en) * | 2016-01-07 | 2016-05-18 | 温州大学 | Acinetobacter and application of acinetobacter in removal of nitrogen and phosphorus from wastewater |
| CN108359617A (en) * | 2017-12-29 | 2018-08-03 | 浙江双良商达环保有限公司 | Acinetobacter calcoaceticus CL05 and its application in the processing of villages and small towns sewage dephosphorization |
| CN108774625A (en) * | 2017-12-29 | 2018-11-09 | 浙江双良商达环保有限公司 | Acinetobacter calcoaceticus CL04 and its application in the processing of villages and small towns sewage dephosphorization |
| CN109022328A (en) * | 2018-09-05 | 2018-12-18 | 海南师范大学 | The application of one plant of polyP bacteria and its Polyphosphate kinase gene in sewage dephosphorization |
| CN109402008A (en) * | 2018-11-15 | 2019-03-01 | 中国农业科学院饲料研究所 | One plant of acinetobacter calcoaceticus TAT1-6A and its application with indoles degradation capability |
| CN110106108A (en) * | 2019-04-25 | 2019-08-09 | 黄山市益天士生物科技有限公司 | Method for improving purifying water body breeding colony speed |
| CN110106097A (en) * | 2019-04-25 | 2019-08-09 | 黄山市益天士生物科技有限公司 | Accelerate the strain enrichment procedure of reparation eutrophication water |
| CN113462595A (en) * | 2021-06-18 | 2021-10-01 | 江南大学 | Efficient phosphorus-accumulating strain obtained by ARTP mutagenesis |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105586294A (en) * | 2016-01-07 | 2016-05-18 | 温州大学 | Acinetobacter and application of acinetobacter in removal of nitrogen and phosphorus from wastewater |
| CN108359617A (en) * | 2017-12-29 | 2018-08-03 | 浙江双良商达环保有限公司 | Acinetobacter calcoaceticus CL05 and its application in the processing of villages and small towns sewage dephosphorization |
| CN108774625A (en) * | 2017-12-29 | 2018-11-09 | 浙江双良商达环保有限公司 | Acinetobacter calcoaceticus CL04 and its application in the processing of villages and small towns sewage dephosphorization |
| CN108359617B (en) * | 2017-12-29 | 2021-03-02 | 浙江双良商达环保有限公司 | Acinetobacter CL05 and application thereof in village and town sewage dephosphorization treatment |
| CN109022328A (en) * | 2018-09-05 | 2018-12-18 | 海南师范大学 | The application of one plant of polyP bacteria and its Polyphosphate kinase gene in sewage dephosphorization |
| CN109022328B (en) * | 2018-09-05 | 2019-11-05 | 海南师范大学 | The application of one plant of polyP bacteria and its Polyphosphate kinase gene in sewage dephosphorization |
| CN109402008A (en) * | 2018-11-15 | 2019-03-01 | 中国农业科学院饲料研究所 | One plant of acinetobacter calcoaceticus TAT1-6A and its application with indoles degradation capability |
| CN110106108A (en) * | 2019-04-25 | 2019-08-09 | 黄山市益天士生物科技有限公司 | Method for improving purifying water body breeding colony speed |
| CN110106097A (en) * | 2019-04-25 | 2019-08-09 | 黄山市益天士生物科技有限公司 | Accelerate the strain enrichment procedure of reparation eutrophication water |
| CN113462595A (en) * | 2021-06-18 | 2021-10-01 | 江南大学 | Efficient phosphorus-accumulating strain obtained by ARTP mutagenesis |
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