CN106244502A - One strain efficient dephosphorization and the pseudomonas of degraded lecithin - Google Patents
One strain efficient dephosphorization and the pseudomonas of degraded lecithin Download PDFInfo
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Abstract
The invention discloses a strain efficient dephosphorization and degraded lecithin function pseudomonas (Pseudomonas sp.) MET69, described bacterial strain JIUYUE in 2016 has been preserved in China typical culture collection center on 12nd, deposit number is CCTCC NO:M2016474, bacterial strain MET69 of the present invention has efficient dephosphorization ability, when with this bacterium and processing low concentration (P≤10mg/L) and high concentration (P about 20mg/L) phosphorus-containing wastewater respectively with the chitosan crosslinked immobilized spherule being prepared as, waste water all can be made to meet " urban wastewater treatment firm pollutant emission standard " (GB18918 2002) one-level B standard (P < 0.5mg/L), compared with reporting bacterial strain, there is clear superiority；Studying simultaneously and also find, bacterial strain MET69 can degrade lecithin, and the preparation for bio-bacterial manure and phosphorus decomposing enzyme preparation provides excellent material, illustrates that this bacterial strain is administered at phosphorus-containing wastewater and has broad application prospects in agricultural production.
Description
Technical field
The invention belongs to environmental microbiology field, be specifically related to a strain efficient dephosphorization and the pseudomonas of degraded lecithin
And the application at aspects such as waste water process, agriculturals.
Background technology
In recent decades, along with China's population increase, industrialized greatly developing, body eutrophication pollution problem is more
Seriously.It is generally believed that phosphorus is the main incitant of body eutrophication, waste water dephosphorization is prevent body eutrophication important
Approach；On the other hand, phosphorus is the main nutrient elements needed for plant growing, and phosphorus element deficiency can have a strong impact on the growth of plant.For
Realizing the sustainable development of phosphor resource, at present in waste water dephosphorization field, " process " of how becoming traditional is " recovery ", by
Gradually become the study hotspot of countries in the world scholar and government pays close attention to object.
Rhodopseudomonas is extensive in distributed in nature, is modal a kind of gram negative bacteria present in soil, its
In some possess the functions such as antibacterial, nitrification and denitrification, Biological control and environmental improvement are of great importance.The most relevant false single
The report of born of the same parents bacterium is concentrated mainly on the aspect such as Biological control and waste water process.Such as patent 201510400702.3(China, Liu Lihe
Deng) report a Pseudomonas aeruginosa strain, can effectively prevent and treat the leaf blight of corn, Causal Organism of Maize Basal Stalk, maize sheath blight, cabbage leaf
The plant diseases such as maize ear rot, asparagus bean rust, Folium Capsici maize ear rot；Patent 201410598044.9(China, Li Qiang etc.) it is found that a strain
Can process the short distance nitration antibacterial of ammonia nitrogen in sewage, be identified as pseudomonas, the total nitrogen in sewage and COD are had certain by it
Removal ability；Patent 200910032681.9(China, brightness Liu etc.) disclose a strain can solve lecithin, promote poplar seedlings growth
Bacillus cercus；At patent 201410665101.0(China, Zhu Xikun etc.) in mention a strain and can degrade different phenol
The pseudomonas of compounds, has good using value in processing the industrial wastewater containing phenol；Patent 201010538869.3
(China, Yuan Hongli etc.) then reports a strain Pseudomonas stutzeri and the application etc. in water body dephosphorized thereof.At present, there is no can
Process phosphorus sewage and have again the pseudomonas report of degraded lecithin function, and published phosphorus sewage water treatment method can not realize
Recycling to phosphorus.
Bacterial strain MET69 of the present invention be a pseudomonas (Pseudomonas sp.), have dephosphorization and lecithin of degrading concurrently
Function.After this bacterial strain, compared with existing report, has a significant advantage in dephosphorization ability, and this bacterial strain makes immobilized spherule, no
Only phosphor-removing effect promotes further, and also is capable of the recovery to phosphorus and recycles.Additionally, the phosphorus in soil has 20%
～50% be to exist with the state of phytic acid, nucleoprotein and lecithin, it is difficult to being absorbed and used by plants, bacterial strain MET69 is to lecithin
Have Degradation, be applied in soil the Phosphorus supply capacity that can significantly improve soil to plant, preparation bio-bacterial manure and
Phosphorus decomposing enzyme preparation has certain application prospect.Therefore, phosphorus waste water is processed by bacterial strain MET69 of the present invention and biological husbantry is sent out
Exhibition is respectively provided with significance.
Summary of the invention
It is an object of the invention to provide a strain efficient dephosphorization and degraded lecithin pseudomonas (Pseudomonas sp.) MET69, and being fixed bead is applied to waste water dephosphorization.
The present invention is achieved through the following technical solutions above-mentioned purpose:
One strain efficient dephosphorization and the pseudomonas of degraded lecithin, Classification And Nomenclature is: pseudomonas (Pseudomonas sp.)
MET69, has been preserved in China typical culture collection center, deposit number: CCTCC NO:M2016474, preservation date:
On JIUYUE 12nd, 2016.
The 16S rDNA sequence of bacterial strain MET69, is shown in sequence table.
The cultural method of above-mentioned pseudomonas MET69: beef-protein medium (peptone 10 g/L, Carnis Bovis seu Bubali cream 3
G/L, NaCl5 g/L, distilled water 1000ml, pH 7.0 ~ 7.2), liquid amount 100 mL(250 mL conical flask), seed liquor is inoculated
The 1% of amount 1%(beef-protein medium volume), cultivate 10 h with 150 r/min at 28 DEG C.
The efficient dephosphorization research of above-mentioned pseudomonas MET69: phosphorus-containing wastewater (PAM culture medium) formula (g/L): sodium citrate
4.0, sodium chloride 0.5, ammonium sulfate 2.5, calcium chloride 0.25, magnesium sulfate 0.25, potassium dihydrogen phosphate 0.0439, maltose 0.01, first
Aniline blue-O 0.025, distilled water 1000ml, pH7.2,121 DEG C of sterilizing 30min.
The research of the immobilized spherule high phosphorus concentration waste water of process of above-mentioned pseudomonas MET69: simulated wastewater formula (g/
L): glucose 0.3, peptone 0.1, sodium acetate 0.15, sodium chloride 0.05, bitter salt 0.15, ammonium chloride 0.18, phosphorus
Acid dihydride potassium (in terms of P: 20:mg/L, 0.0878), distilled water 1000mL, pH7.0,115 DEG C of sterilizing 25min.
The research that lecithin is degraded by above-mentioned pseudomonas MET69: basal medium: glucose 10.0 g, (NH4)2SO40.5 g, MgSO4·7H2O 0.3 g, NaCl0.3g, KCl0.3 g, FeSO4·7H2O 0.03 g, MnSO4·4H2O
0.03 g, agar 20 g, distilled water 1000 mL, pH are natural, 115 DEG C of sterilizing 25min.50～60 DEG C it are cooled to after sterilizing.Will
Taking egg yolk after one raw egg surface sterilization and pour in sterilizing triangular flask, add 50ml sterilized water, mixing is added by addition 10%
Enter in basal medium, obtain lecithin medium.
Comparing with existing patented strain, bacterial strain of the present invention has the advantage that
(1), when processing low concentration wastewater (P≤10mg/L) with bacterial strain MET69 of the present invention, waste water can be made in 24h to meet, and " cities and towns are dirty
Water treatment plant's pollutant emission standard " (GB18918-2002) one-level B standard (P < 0.5mg/L)；With bacterial strain MET69 of the present invention with
When chitosan crosslinked immobilized spherule processes high phosphorus concentration waste water (P about 20 mg/L), 48h tp removal rate, up to 98.54%, is located
" urban wastewater treatment firm pollutant emission standard " (GB18918-2002) one-level B standard (P < 0.5mg/L) can be met after reason, say
The dephosphorization capacity of bright bacterial strain MET69 is totally preferable, and good environmental adaptability, and in industrialization, using value is big.
(2) bacterial strain MET69 of the present invention has preferable degradation capability to lecithin, it is possible to be unique phosphorus source with lecithin
Culture medium on form significant phosphorus decomposing circle, MET69 microbial inoculum is imposed in soil, it is possible to the rapid available phosphorus significantly improving soil contains
Amount, is good soil conditioner.
Accompanying drawing explanation
Fig. 1 is many dephosphorization hydrochlorate Color of MET69 bacterial strain.
Fig. 2 is MET69 growth curve in beef extract-peptone fluid medium.
Fig. 3 is MET69 bacterial strain cultivation form on beef extract-peptone flat board.
Fig. 4 is the 16S rDNA gene order phylogenetic tree of MET69 bacterial strain.
Fig. 5 is that the chitosan-immobilized bead of MET69 processes high phosphorus concentration waste water effect.
Fig. 6 is the MET69 degradation effect to lecithin.
Detailed description of the invention
The following is the specific embodiment of the present invention, technical scheme is described further, but the present invention's is interior
Hold the scope being not limited solely to described in embodiment, every change without departing substantially from present inventive concept or equivalent replacement and be included in this
Within the protection domain of invention.
The nutrient media components related in case study on implementation:
PAM-TBO culture medium (g/L): sodium citrate 4.0, sodium chloride 0.5, ammonium sulfate 2.5, calcium chloride 0.25, magnesium sulfate 0.25,
Potassium dihydrogen phosphate 0.0439, maltose 0.01, toluidine blue-O 0.025, distilled water 1000ml, pH7.2, PAM-TBO culture medium
Qualitative screening for dephosphorization bacterial.
Rich phosphorus culture medium (g/L): sodium acetate 5, ammonium chloride 2, potassium dihydrogen phosphate 0.0439, bitter salt 0.5, chlorine
Changing calcium 0.2, distilled water 1000ml, pH7.0 ~ 7.2, enrichment medium is for the activation of dephosphorization bacterial.
Beef-protein medium (g/L): peptone 10, Carnis Bovis seu Bubali cream 3, NaCl 5, distilled water 1000ml, pH7.0 ~
7.2, beef-protein medium is used for isolated and purified, preservation and the domestication of bacterial strain.
Simulated wastewater formula (g/L): glucose 0.3, peptone 0.1, sodium acetate 0.15, sodium chloride 0.05, seven hydration sulfur
Acid magnesium 0.15, ammonium chloride 0.18, potassium dihydrogen phosphate (in terms of P: 10mg/L, 0.0439；In terms of P: 20:mg/L, 0.0878), steam
Distilled water 1000mL, pH7.0.
Lecithin medium: basal medium (g/L): glucose 10.0, (NH4)2SO40.5, MgSO4·7H2O
0.3, NaCl 0.3, KCl 0.3, FeSO4·7H2O 0.03, MnSO4·4H2O 0.03, agar 20, distilled water 1000 mL
, pH is natural, 115 DEG C of sterilizing 25min.50～60 DEG C it are cooled to after basal medium sterilizing.After a raw egg surface sterilization
Taking egg yolk and pour in sterilizing triangular flask, add 50ml sterilized water, mixing adds in basal medium by addition 10%, to obtain final product
Lecithin medium.
Embodiment 1 pseudomonas (Pseudomonas sp.) screening of MET69
VasviChaudhry and Chandra ShekharNautiyal is once at the " Bioresource of 17 phases of volume 102 in 2011
Technology " on report a kind of method of high flux screening dephosphorization bacterial, utilize the toluidine blue can be to removing in bacterial strain more
The characteristic of phosphate particle dyeing, can be with Rapid identification dephosphorization bacterial.Bacterial strain of the present invention is i.e. obtained by the method screening, specifically grasps
Make step:
1. Yangxin County, Huangshi, the Hubei Province chemical plant wastewater sample collected is mixed by the dilution proportion of 1:9 with sterilized water, institute
Obtaining suspension concentration is 10-1；
The most again this bacteria suspension 10 doubling dilution is become variable concentrations, be respectively coated in rich phosphorus culture medium, be inverted for 28 DEG C and cultivate
48h；
3. the bacterium colony that picking morphological characteristic is different carries out purification of ruling, and is numbered purification bacterial strain, retains standby；
4. above-mentioned bacterial strains activation being obtained seed liquor, on 96 orifice plates, every hole adds 0.18ml liquid PAM-TBO culture medium respectively, and
Inoculating 20 μ L bacterium solution, every three the Kong Weiyi groups of file, the most each bacterial strain does 3 repetitions, not inoculate the process of bacterium sample as right
According to；
5. with sealed membrane by 96 pore plate by sealing, just putting 28 DEG C and cultivating 2 ~ 3d, observing discoloration in orifice plate, result is shown in Fig. 1.Fade
More clearly show that bacterial strain dephosphorization ability is the strongest, the obvious bacterial strain that fades is carried out glycerol method preservation.
By above-mentioned screening technique, it is thus achieved that 1 strain can make the bacterial strain that toluidine blue culture medium is significantly faded, numbered
MET69, i.e. this patent bacterial strain.It is seeded to by 1% inoculum concentration (accounting for the 1% of beef-protein medium volume) after MET69 activation
Beef extract-peptone fluid medium, growth curve is shown in Fig. 2.The logarithmic (log) phase of MET69 is about at 10 ~ 28h as seen from the figure, now
MET69 activity is the strongest, and dephosphorization efficiency is higher.
The qualification of embodiment 2 bacterial strain MET69
(1) morphological characteristic and physiological and biochemical property
The form of bacterial strain MET69 and physiological and biochemical property: 28 DEG C of constant temperature culture 48h on beef extract-peptone flat board, bacterium colony is 2
~ 3mm is circular, and the smooth of the edge, in faint yellow, opaque, concrete colonial morphology is shown in Fig. 3.With reference to " primary Jie Shi Bacteria Identification handbook ",
Obtaining this bacterial strain and also have the following characteristics that Gram-negative, thalline is shaft-like, is formed without spore and pod membrane, and amphimicrobian, without whip
Hair, catalase positive, amylase reaction negative, C.I. 13020. is negative, V-P reaction negative.
(2) 16S rDNA Molecular Identification
Carry out the PCR amplification in 16S rDNA V3 district with bacterial strain DNA for template, primer sequence is: 27F
(AGAGTTTGATCCTGGCTCAG) and 1492R(GGTTACCTTGTTACGACTT)；
PCR system sets up (50 μ L): Template DNA 1 μ L, Forward Primer (10uM) 1 μ L, Reverse
Primer (10uM) 1 μ L, Taq-Plus 1 μ L, Taq-Plus Buffer 5 μ L, dNTPs (2mM) 5 μ L, sterilized water 36 μ L.
PCR response parameter is: 95 DEG C of denaturations 5min, 94 DEG C of degeneration 30s, 55 DEG C of renaturation 45s, and 72 DEG C extend 90s, totally 30
Individual circulation, 72 DEG C extend 10min the most again.
After PCR terminates, take 5 μ LPCR products and carry out 0.8% agarose gel electrophoresis detection, purpose clip size about 1.5kb.
Deliver to Wuhan after PCR primer being reclaimed hold up Kechuang neoplasm Science and Technology Ltd. and check order.Checked order a length of 1566bp of row.
Measured sequence carries out on NCBI BLAST comparison, and result display sequence and bacterial strain MET69 similarity reach 99%
Be Rhodopseudomonas.Download the 16S rDNA sequence that dependency is higher, with the phylogeny of MEGA software building MET69
Tree, sees Fig. 4.
In conjunction with strain morphology, physiological and biochemical property and 16S rDNA sequence alignment analysis, finally determine that bacterial strain MET69 is one
Pseudomonas (Pseudomonas sp.)。
Embodiment 3 bacterial strain of the present invention phosphorus removal property is studied
In order to evaluate the phosphorus removal property of bacterial strain of the present invention, bacterial strain MET69 is respectively 5,10,15,20 mg/L at phosphorus concentration and contains
Dephosphorizing rate in phosphorus waste water (PAM culture medium) is measured and compares.
By bacterial strain MET69 activation culture to OD600Being 1.0, the inoculum concentration (for the volume 10% of PAM culture medium) by 10% turns
Be connected in the PAM culture medium (in terms of P: 5,10,15,20 mg/L) of 200 mL, in 28 DEG C, cultivate under the conditions of 150 r/min, every
12,24,36,48,60(h) sampling, after 10000 r/min are centrifuged 10 min, molybdenum antimony resistance colorimetric method measures the phosphorus concentration of supernatant
(GB 11893-89), and calculate dephosphorizing rate (the results are shown in Table 1).
Dephosphorizing rate (%)=(XSample introduction-XGo out sample)/XSample introduction× 100%
XSample introduction: access the total phosphorus concentration of the enrichment culture liquid before bacterial strain
XGo out sample: access the total phosphorus concentration of the enrichment culture liquid after bacterial strain
Table 1 MET69 processes different phosphate concentration waste water effectiveness comparison
Result shows, bacterial strain of the present invention can efficiently process low concentration (P≤10mg/L) phosphorus-containing wastewater, can meet after process in 24h
" urban wastewater treatment firm pollutant emission standard " (GB18918-2002) one-level B standard (P < 0.5mg/L), phosphor-removing effect shows
Write, but when processing the phosphorus-containing wastewater of higher concentration (10 < P≤20mg/L), still can not meet discharge standard.In order to improve bacterial strain
The phosphorus removal property of MET69 and cycle performance, to bacterial strain being fixed treatment research.
Embodiment 4 bacterial strain MET69 immobilized spherule height phosphorus concentration waste water (P about 20 mg/L) processes
The preparation of immobilization blank bead: 2.5% chitosan gum liquid solution is instilled condensation water (20% methanol and 30% with No. 8 syringe needles
NaOH) in, obtaining hollow globular chitosan ball after acting on 2 h, taking-up is washed with deionized, standby.
The preparation of immobilization embedded bacterium bead: the immobilization blank bead prepared is put into wet bacterium and sterile saline
The bacteria suspension that mass volume ratio (g:ml) is 2:100 in, adsorb 2 h, take out chitosan bead and also wash with sterile saline
The most standby.
Weigh 2g immobilization blank bead, immobilization embedded bacterium bead respectively, be then respectively put into equipped with 200 ml phosphorus dense
Degree is in 500 ml conical flasks of 20 mg/L waste water, 28 DEG C, 150 r/min shaking tables process, at regular intervals 0,12,
24,36,48,72(h) take waste water 5 ml, in centrifuge, 10000 r/min are centrifuged 10 min, take supernatant, divide according to ammonium molybdate
Light photometry (GB11893-89) measures wherein total phosphorus content, calculates dephosphorizing rate.
Experimental result is shown in accompanying drawing 5, as it can be seen, the immobilization embedded bacterium bead high phosphorus concentration waste water of process is when 48 h, removes
Phosphorus effect reaches optimal, and dephosphorizing rate is 98.54%, and now the phosphorus concentration in system is 0.292 mg/L.
Test result indicate that, process high phosphorus concentration waste water (P about 20 with the crosslinking bead of pseudomonas MET69 with chitosan
Mg/L), it is possible to make it meet " urban wastewater treatment firm pollutant emission standard " (GB18918-2002) one-level B in 48 h
Standard (P < 0.5mg/L).After illustrating that bacterial strain MET69 is immobilized, not only dephosphorization ability improves further, also achieves phosphorus element
Recovery and recycle.
The embodiment 5 bacterial strain MET69 Degradation to lecithin
The Degradation of lecithin is used phosphorus decomposing circle method to verify, by MET69 at beef extract-peptone liquid culture by MET69
After base activates 10h, at the flat lining out of lecithin (point sample), it is inverted after cultivating 48h at 37 DEG C, it is seen that the bacterium colony week of MET69
Cross now significantly phosphorus decomposing circle (phosphorus decomposing circle refers on the solid medium containing organophosphor, periphery of bacterial colonies produce because of organophosphor
The transparent circle degraded and formed), result is shown in accompanying drawing 6.MET69 can on flat board that lecithin is unique phosphorus source growth shape
Become transparent phosphorus decomposing circle, illustrate that MET69 has preferable Degradation to lecithin.
Embodiment 6 MET69 impact on content of soil available phosphor
Take the unmanured normal soil of university's periphery as reference, the checking MET69 impact on soil quick-effective phosphor.Soil quick-effective phosphor
Also referred to as soil available phosphorus, including water-soluble phosphorus and weak acid dissolubility phosphorus, rapid available phosphorus is easily absorbed and used by plants, therefore its content is to sentence
One important indicator of disconnected soil Phosphorus supply capacity.The NaHCO of the present embodiment 0.5mol/L3(pH8.5) it is that digestion agent processes soil
Earth, the phosphorus content of leachate uses molybdenum antimony resistance colorimetric method (GB 11893-89) to measure.
After MET69 is activated 10h in beef extract-peptone liquid culture, collect thalline and by sterile water wash 1 time, then
OD is adjusted with sterilized water600To 1.0, now bacteria concentration about 3 × 108cfu/mL.Every 100g soil sample adds the MET69 bacterium solution of 10mL also
Stirring, be placed under natural environment and in 0h, 24h, 48h, 72h are separately sampled.
Weigh air-dried soil sample 5.00g by 1mm sieve aperture to be placed in 250ml triangular flask, add one little spoonful of non-phosphorus active carbon
NaHCO with 0.5mol/L3(pH8.5) lixiviating solution 100ml, stoppers bottle stopper in acutely shaking 30min, immediately with being dried after taking-up
Funnel and without phosphorus filter paper filtering, filtrate is with another triangular flask access.Take 10mL filtrate and measure phosphorus concentration with molybdenum antimony resistance colorimetric method,
Substitute into formula and calculate corresponding content of soil available phosphor, the results are shown in Table 2.
Content of soil available phosphor (mg/kg)=
Note:
Table 2 MET69 impact on soil available phosphorus content in 72h
As shown in Table 2, relatively low for examination soil Phosphorus supply capacity originally, but after MET69 microbial inoculum is applied to soil 72h, for examination
Content of available phosphorus in soil significantly improves, and Phosphorus supply capacity is medium level on the upper side, illustrates that MET69 is good soil conditioner,
Can promote that soil supplements phosphorus element for plant in time, use reducing chemical fertilizer, reduce agricultural cost, it is achieved to soil ring
The biological restoration in border is significant.
Sequence table
<110>Hubei University
<120>one strain efficient dephosphorizations and the pseudomonas of degraded lecithin
<160>3
<210>1
<211>1566
<212>DNA
<213>pseudomonas (Pseudomonas sp.)
<400>1
ggcctgcagg tcgacgattg gttaccttgt tacgacttca ccccagtcat gaatcactcc 60
gtggtaaccg tcccccttgc ggttagacta gctacttctg gagcaaccca ctcccatggt 120
gtgacgggcg gtgtgtacaa ggcccgggaa cgtattcacc gtgacattct gattcacgat 180
tactagcgat tccgacttca cgcagtcgag ttgcagactg cgatccggac tacgatcggt 240
tttatgggat tagctccacc tcgcggcttg gcaacccttt gtaccgacca ttgtagcacg 300
tgtgtagccc tggccgtaag ggccatgatg acttgacgtc atccccacct tcctccggtt 360
tgtcaccggc agtctcctta gagtgcccac ccgaggtgct ggtaactaag gacaagggtt 420
gcgctcgtta cgggacttaa cccaacatct cacgacacga gctgacgaca gccatgcagc 480
acctgtgtct gagttcccga aggcaccaat ccatctctgg aaagttctca gcatgtcaag 540
gccaggtaag gttcttcgcg ttgcttcgaa ttaaaccaca tgctccaccg cttgtgcggg 600
cccccgtcaa ttcatttgag ttttaacctt gcggccgtac tccccaggcg gtcgacttat 660
cgcgttagct gcgccactaa gatctcaagg atcccaacgg ctagtcgaca tcgtttacgg 720
cgtggactac cagggtatct aatcctgttt gctccccacg ctttcgcacc tcagtgtcag 780
tatcagtcca ggtggtcgcc ttcgccactg gtgttccttc ctatatctac gcatttcacc 840
gctacacagg aaattccacc accctctacc gtactctagc tcagtagttt tggatgcagt 900
tcccaggttg agcccgggga tttcacatcc aacttgctga accacctacg cgcgctttac 960
gcccagtaat tccgattaac gcttgcaccc ttcgtattac cgcggctgct ggcacgaagt 1020
tagccggtgc ttattctgtt ggtaacgtca aaacagcaag gtattaactt actgcccttc 1080
ctcccaactt aaagtgcttt acaatccgaa gaccttcttc acacacgcgg catggctgga 1140
tcaggccttc gcccattgtc caatattctc cactgctgcc tcccgtagga gtctggaccg 1200
tgtctcagtt ccagtgtgac tgatcatcct ctcagaccag ttacggatcg tcgccttggt 1260
aggcctttac cccaccaact agctaatccg acctaggctc atctgatagc gtgaggtccg 1320
aagatccccc actttctccc tcaggacgta tgcggtatta gcgcccgttt ccggacgtta 1380
tcccccacta ccaggcagat tcctaggcat tactcacccg tccgccgctg aatccaggag 1440
caagctccct tcatccgctc gacttgcatg tgttaggcct gccgccagcg ttcaatctga 1500
gccaggatca aactctaatc tctagaggat ccccgggtac cgagctcgaa ttcgtaatca 1560
gttttc 1566
<210> 2
<211> 20
<212> DNA
<213>artificial sequence
<400> 2
agagtttgat cctggctcag 20
<210> 3
<211> 19
<212> DNA
<213>artificial sequence
<400> 3
ggttaccttg ttacgactt 19
Claims (3)
1. a strain efficient dephosphorization and the pseudomonas of degraded lecithin, it is characterised in that: this pseudomonas isPseudomonas sp.MET69, within 2016, JIUYUE has been preserved in China typical culture collection center on 12nd, and deposit number is CCTCC NO:M
2016474。
2. a strain efficient dephosphorization and the pseudomonas of degraded lecithin, it is characterised in that: this pseudomonasPseudomonas sp.MET69 and immobilized spherule thereof can process phosphorus-containing wastewater.
3. a strain efficient dephosphorization and the pseudomonas of degraded lecithin, it is characterised in that: this pseudomonasPseudomonas sp.MET69 has Degradation to lecithin, can be applicable to bio-bacterial manure and the preparation of phosphorus decomposing enzyme preparation.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108546658A (en) * | 2018-04-10 | 2018-09-18 | 暨南大学 | One plant of phosphorus-solubilizing bacteria and its compound microbial inoculum and application with DEHP degradation bacterias |
| CN108795799A (en) * | 2018-05-31 | 2018-11-13 | 山东省科学院生态研究所 | One plant of multi-functional arthrobacterium Fp64 and its application for controlling phosphorus levels |
| CN116891809A (en) * | 2022-12-06 | 2023-10-17 | 浙江大学 | Pseudomonas asiatica and microbial agent and application thereof |
Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1519309A (en) * | 2003-05-22 | 2004-08-11 | 郑爱萍 | Strain of pseudomonas for promoting growth of paddy as well as preventing and curing sheath blight of rice |
| CN101698881A (en) * | 2007-12-18 | 2010-04-28 | 曲奕 | Lecithase test medium |
| CN101709217A (en) * | 2009-12-03 | 2010-05-19 | 中国农业大学 | Pseudomonas fluorescens CKD18 and application thereof |
| CN101935622A (en) * | 2010-02-05 | 2011-01-05 | 北京大学 | Pseudomonas strain and application thereof |
| CN102586163A (en) * | 2012-02-15 | 2012-07-18 | 南京大学 | Polyphosphate transgenic pseudomonas putida engineering bacterium as well as construction method and application thereof |
| CN102776145A (en) * | 2012-07-31 | 2012-11-14 | 山东大学 | Denitrifying polyphosphate accumulation bacterium and application of same in sewage treatment |
| CN102864098A (en) * | 2012-04-26 | 2013-01-09 | 哈尔滨工业大学宜兴环保研究院 | Denitrification phosphorus removal bacterium H-hrb02 as well as screening method and application thereof |
| 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 |
| CN103451116A (en) * | 2012-06-04 | 2013-12-18 | 华中农业大学 | Pseudomonas fluorescens Y13, and preparation method and applications thereof |
| CN105754903A (en) * | 2016-04-15 | 2016-07-13 | 中国水产科学研究院渔业机械仪器研究所 | Method for cultivating photosynthetic bacteria in Rhodopseudomonas on large scale |
| CN106422149A (en) * | 2016-09-23 | 2017-02-22 | 杭州富阳佳畅机械有限公司 | Biological preparation of degrading organophosphorus pesticide residues |
-
2016
- 2016-09-27 CN CN201610853898.6A patent/CN106244502B/en active Active
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1519309A (en) * | 2003-05-22 | 2004-08-11 | 郑爱萍 | Strain of pseudomonas for promoting growth of paddy as well as preventing and curing sheath blight of rice |
| CN101698881A (en) * | 2007-12-18 | 2010-04-28 | 曲奕 | Lecithase test medium |
| CN101709217A (en) * | 2009-12-03 | 2010-05-19 | 中国农业大学 | Pseudomonas fluorescens CKD18 and application thereof |
| CN101935622A (en) * | 2010-02-05 | 2011-01-05 | 北京大学 | Pseudomonas strain and application thereof |
| CN102586163A (en) * | 2012-02-15 | 2012-07-18 | 南京大学 | Polyphosphate transgenic pseudomonas putida engineering bacterium as well as construction method and application thereof |
| CN102864098A (en) * | 2012-04-26 | 2013-01-09 | 哈尔滨工业大学宜兴环保研究院 | Denitrification phosphorus removal bacterium H-hrb02 as well as screening method and application thereof |
| CN103451116A (en) * | 2012-06-04 | 2013-12-18 | 华中农业大学 | Pseudomonas fluorescens Y13, and preparation method and applications thereof |
| CN102776145A (en) * | 2012-07-31 | 2012-11-14 | 山东大学 | Denitrifying polyphosphate accumulation bacterium and application of same in sewage treatment |
| 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 |
| CN105754903A (en) * | 2016-04-15 | 2016-07-13 | 中国水产科学研究院渔业机械仪器研究所 | Method for cultivating photosynthetic bacteria in Rhodopseudomonas on large scale |
| CN106422149A (en) * | 2016-09-23 | 2017-02-22 | 杭州富阳佳畅机械有限公司 | Biological preparation of degrading organophosphorus pesticide residues |
Non-Patent Citations (2)
| Title |
|---|
| 刘诚等: "多功能菌株假单胞菌的溶磷和解磷效果及其应用", 《湖北大学学报（自然科学版）》 * |
| 陈倩颖: "解有机磷细菌的分离鉴定及其解磷特性研究", 《安徽农业大学》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108546658A (en) * | 2018-04-10 | 2018-09-18 | 暨南大学 | One plant of phosphorus-solubilizing bacteria and its compound microbial inoculum and application with DEHP degradation bacterias |
| CN108546658B (en) * | 2018-04-10 | 2020-09-04 | 暨南大学 | Phosphorus-dissolving bacterium and compound bacterium agent of phosphorus-dissolving bacterium and DEHP degrading bacterium and application of phosphorus-dissolving bacterium and compound bacterium agent |
| CN108795799A (en) * | 2018-05-31 | 2018-11-13 | 山东省科学院生态研究所 | One plant of multi-functional arthrobacterium Fp64 and its application for controlling phosphorus levels |
| CN116891809A (en) * | 2022-12-06 | 2023-10-17 | 浙江大学 | Pseudomonas asiatica and microbial agent and application thereof |
| CN116891809B (en) * | 2022-12-06 | 2023-12-19 | 浙江大学 | Pseudomonas asiatica and microbial agent and application thereof |
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| CN106244502B (en) | 2019-06-18 |
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