CN100422312C - Chlorine resistant strain V430 and its screening method - Google Patents
Chlorine resistant strain V430 and its screening method Download PDFInfo
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- CN100422312C CN100422312C CNB2006100181337A CN200610018133A CN100422312C CN 100422312 C CN100422312 C CN 100422312C CN B2006100181337 A CNB2006100181337 A CN B2006100181337A CN 200610018133 A CN200610018133 A CN 200610018133A CN 100422312 C CN100422312 C CN 100422312C
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Abstract
The present invention relates to a kind of chlorine tolerant microbe and its screening process. The chlorine tolerant microbe strain is Staphylococcus saprophyticus V430 in the preservation number of CCTCC No. M205145. The Staphylococcus saprophyticus V430 has circular opaque colony shape with lug and smooth edges; spherical single, paired or piled cells of d in 0.6-0.9 micron without flagellum; no motion, no endogeneous spore, gram positive and anaerobic property; and optimal growth pH value of 6.0-7.5 and temperature of 25-45 deg.c. It can survive in high chlorine ion concentration environment and degrade high concentration organic pollutant effectively.
Description
Technical field
The present invention relates to a kind of anti-chlorine microorganism and screening method thereof.
Background technology
The high salinity organic wastewater with difficult degradation thereby has become the worldwide technological puzzle of environmental protection water treatment field as waste water such as oil production, chemical industry, pharmacy.And salt acid system turmeric saponin factory effluent is representative in high density saliferous hardly degraded organic substance waste water.On the one hand, this class waste water complicated component, the organism of difficult degradation is many, and the biodegradability of waste water is poor; On the other hand, the salts contg height in the waste water, chlorine ion concentration is big, and chlorine ion concentration can reach 20000mg/L in its composite waste.At present, salt-containing organic wastewater adopts abiotic method and biological method, abiotic method is owing to its processing costs height, treatment effect are bad, enterprise can't accept, biochemical process is the at first method in the water technology always, the main two sections contact oxidation methods of A-B that adopt, traditional activated sludge process, the SBR method, biological filter, anaerobic filter etc., it is waste water below 3% that but above-mentioned biological process can only be handled saltiness mostly, and high-salt wastewater (saltiness 5%, even 20%) is difficult to handle, therefore need special microorganism.In recent years, reported that abroad relevant seed selection salt tolerant (NaCl) bacterium handles the report of high salt organic waste water, and to this class high-enriched organics content of saponin waste water, high salt (CaCl2), contain the complicated waste water of hard-degraded substance (saponin), also belong to the weak field of research so far.
Summary of the invention
The anti-chlorine strain V 430 and the screening method thereof that the object of the present invention is to provide a strain to handle to contain high-concentration chlorine ion wastewater.
Anti-chlorine strain V 430 provided by the present invention, be Staphylococcus saprophyticus (Staphylococcus saprophyticus) V430CCTCC NO.M205145, Chinese typical culture collection center (being called for short CCTCC), preserving number: CCTCC NO.M205145 have been preserved in on December 9th, 2005.
The colonial morphology of described Staphylococcus saprophyticus (Staphylococcus saprophyticus) V430 CCTCC NO.M205145: bacterium colony is rounded, children's oyster white in age, aged safran is opaque; Bacterium colony projection, edge-smoothing; Cell spheroid, d=0.6～single or paired, the in heaps appearance of 0.9 μ m, atrichia; Do not move no endogenous spore, Gram-positive, anaerobism; The suitableeest growth pH value: 6.0～7.5, growth temperature: 25-45 ℃; Main biochemical character: the catalase feminine gender, oxidase negative, glucose produce not aerogenesis of acid; Unhydrolysed casein, gelatin and starch; Can degrade fructose, seminose; Can reductive NO 3
-
The screening method of one chlorine-resisting strain V430,
1), zoogleic fragmentation:
Get the anaerobic activated sludge in the saponin waste water treatment system, press anaerobic activated sludge: sterilized water=2g: the proportioning of 100ml, add in the good triangular flask that sterilized water is housed of sterilization in advance, and adding weight is the trisodium phosphate of sterilized water 0.01%, the shaking table vibration, zoogloea is smashed, got mud mixture, standby;
2), the separation of anti-chlorine ion dominant bacteria:
Utilize the above-mentioned mud mixture of aseptic pipette, extract, press mud mixture: isolation medium=0.5ml: the proportioning of 4.5ml, add isolation medium, 25-45 ℃ of anaerobism constant temperature culture 5-7 days treats that the test tube substratum becomes muddy, illustrates that bacterium grows, dull and stereotyped then dilution coating, picking has single bacterium of notable difference on colony characteristics, until obtaining single bacterial strain, and preserve in the slant medium ramp;
3), the screening of anti-chlorine ion dominant bacteria:
The above-mentioned isolated single bacterium colony of picking is inoculated in respectively in the screening culture medium, 25-45 ℃ of anaerobism constant temperature culture 5-7 days, and the muddy situation of observing bacteria suspension becomes the muddy high chlorine resistant ionic bacterial strain that filters out that is; Through screening, obtain the bacterial strain that a strain is numbered strain V 430 at last, i.e. anti-chlorine strain V 430.
Isolation medium: peptone 10g; Extractum carnis 5g; Sodium-chlor; 5g; Distilled water 1000ml.
Screening culture medium: MgSO47H
2O, 0.2g/L, KH
2PO
40.5g/L, K
2HPO
41.5g/L, NH
4Cl 0.5/L, CaCl
2135g/L, the 1ml trace element solution dissolves in 1L saponin waste water (COD concentration is 1000mg/L), pH=7.5;
Trace element solution: FeCl
36H
2O 2g, CoCl
22g, MnCl
20.5g, CuCl
20.03g, ZnCl
20.05g, NiCl
2.6H
2O 0.05g, EDTA 1g, pH=7.0.
Enlarged culturing base: peptone 10g; Extractum carnis 5g; Sodium-chlor; 5g; Distilled water 1000ml; CaCl
218.0g/L.
SC substratum: peptone 10g; Extractum carnis 5g; Sodium-chlor; 5g; Distilled water 1000ml; CaCl
2Add-on is 12.6g-140g.
Slant medium: peptone 10g; Extractum carnis 5g; Sodium-chlor; 5g; Potassium primary phosphate 2g; Ammonium chloride 1g; Agar 20g; Distilled water 1000ml; CaCl
218g/L, pH=7.5.
Above substratum is standby after respectively at 121 ℃ of autoclaving 30min.
The treatment process that a described staphylococcus saprophyticus (Staphylococcus saprophyticus) V430 is applied to contain high-concentration chlorine ion wastewater is: the single bacterium colony of picking Staphylococcus saprophyticus (Staphylococcus saprophyticus) V430, in the enlarged culturing base, cultivated 20-24 hour, by Staphylococcus saprophyticus (Staphylococcussaprophyticus) V430 after cultivating: the volume ratio that contains high-concentration chlorine ion wastewater (as mixing saponin waste water) is (1-4): 50 get Staphylococcus saprophyticus (Staphylococcus saprophyticus) V430 is inoculated in and contains in the high-concentration chlorine ion wastewater (as mixing saponin waste water), 25-45 ℃ of constant temperature anaerobism cultivated, the pH value is 7.0-8.0, reacts 3-5 days.
Described enlarged culturing base is: peptone 10g; Extractum carnis 5g; Distilled water 1000ml; CaCl
218.0g/L, pH=7.5.
Separate, screen a strain has the certain growth ability under the high-concentration chlorine ion condition new bacterial strain in the active sludge of the present invention from the saponin waste water treatment system.And its anti-chlorine ability studied, the result shows that under the high-concentration chlorine ion condition, the anti-chlorine ability of anti-chlorine strain V 430 is good, scope is wide, can be at [Cl
-] have energy for growth preferably during for 3000-75000mg/L.If it is induced bacterium as adding in the biosystem of the organic waste water of handling high salinity, the microorganism that can overcome general active sludge is subjected to the inhibition of high chloride ion, activity is low, processing efficiency is not high shortcoming.Because in high chloride ion (20000mg/L) environment, thereby being subjected to the effect that chloride permeability presses, the microorganism of traditional technology makes that plasma membrane is impaired, metabolic function descends reduces treatment effect, by contrast, if directly add the high chlorine resistant ion in the biochemical system and have the bacterial strain of good degradation property, just can overcome the shortcoming that above-mentioned traditional technology is handled the high-concentration chlorine ion organic wastewater with difficult degradation thereby.
To contain high salt (CaCl
2The high concentrated organic wastewater of difficult degradation salt)---saponin waste water is an example, and the present invention has studied the effect of its degradable organic pollutant.Found that this bacterial strain can effectively remove the ability of COD in the saponin waste water, the COD that can remove in the waste water within 72 hours is about 50-60%.
But the new bacterial strain that the present invention obtains not only can contain existence under the high-concentration chlorine ion environment, but also efficient degradation high concentration organic contaminant.
Description of drawings
Fig. 1-1a is a strain V 430 bacterial strain Electronic Speculum picture
Fig. 1-1b is a strain V 430 bacterial strain Electronic Speculum picture
Fig. 1-1c is a strain V 430 bacterial strain Electronic Speculum picture
Fig. 1-2 is strain V 430 growing state figure on the SC substratum of the different chlorine ion concentrations of liquid
Fig. 1-the 3rd, and strain V 430 pcr amplification agarose electrophoresis figure (1: nuclear DNA, 2: amplified production, M:Marker)
Fig. 1-4a is the following degraded efficiency diagram of strain V 430 under high chloride ion, high COD concentration
Fig. 1-4b is the following degraded efficiency diagram of strain V 430 under high chloride ion, high COD concentration
Fig. 1-4c is the following degraded efficiency diagram of strain V 430 under high chloride ion, high COD concentration
Fig. 1-4d is the following degraded efficiency diagram of strain V 430 under high chloride ion, high COD concentration
Embodiment
One, the screening method of a chlorine-resisting strain V430:
(1), material is prepared:
1, bacterium source and experiment waste water:
(1) bacterium source: the active sludge in the biochemical system of processing saponin waste water;
(2) experiment waste water (promptly mixing saponin waste water): this experiment waste water is taken from the female Home Co., Ltd in Shiyan City side, Hubei Province diosgenin wastewater; After interior electrolysis and CaO allotment, specific targets are: pH=7.5-8.2, COD=4000-17000mg/L, Cl
-=8000-30000mg/L, NH
4-N 140-300mg/L.
2, substratum:
Separate (fully) substratum: peptone 10g; Extractum carnis 5g; Sodium-chlor; 5g; Distilled water 1000ml.
Screening culture medium: MgSO47H
2O, 0.2g/L, KH
2PO
40.5g/L, K
2HPO
41.5g/L, NH
4Cl 0.5/L, CaCl
2135g/L, the 1ml trace element solution dissolves in 1L saponin waste water (COD concentration is 1000mg/L), pH=7.5;
Trace element solution: FeCl
36H
2O 2g, CoCl
22g, MnCl
20.5g, CuCl
20.03g, ZnCl
20.05g, NiCl
2.6H
2O 0.05g, EDTA 1g, pH=7.0.
Enlarged culturing base: peptone 10g; Extractum carnis 5g; Sodium-chlor; 5g; Distilled water 1000ml; CaCl
218.0g/L.
SC substratum: peptone 10g; Extractum carnis 5g; Sodium-chlor; 5g; Distilled water 1000ml; CaCl
2Add-on is 12.6g-140g; Work as CaCl
2When add-on is 12.6g, [Cl in the SC substratum
-]=10000mg/L.
Slant medium: peptone 10g; Extractum carnis 5g; Sodium-chlor; 5g; Potassium primary phosphate 2g; Ammonium chloride 1g; Agar 20g; Distilled water 1000ml; CaCl
218g/L, pH=7.5.
Above substratum is standby after respectively at 121 ℃ of autoclaving 30min.
3, laboratory apparatus and equipment:
State China SHA-B constant temperature oscillator,
SHH150G illumination bio-incubator,
Horizontal pressure steam sterilizer,
WMX-1 type microwave seal is cleared up the COD tacheometer,
The 722S spectrophotometer,
Opticmicroscope,
PH meter,
Bechtop,
Air dry oven,
Ultramicrotome,
The H-7000FA of Hitachi projection microscope,
PTC200 type PCR instrument,
Electrophoresis apparatus and electrophoresis chamber,
Gel ultraviolet visualizer etc.
(2), the separation of bacterial strain and screening:
1, zoogleic fragmentation:
Get the anaerobic activated sludge in the 2g saponin waste water treatment system, add in the triangular flask of the 250ml that the 100ml sterilized water is housed that sterilization is good in advance, and adding weight is the trisodium phosphate of sterilized water 0.01%, the shaking table vibration, zoogloea is smashed, got mud mixture, standby.
2, the separation of anti-chlorine ion dominant bacteria:
Utilize the above-mentioned mud mixture of aseptic pipette, extract 0.5ml, add isolation medium 4.5ml, 35 ℃ of anaerobism constant temperature culture 5-7 days, treat that the test tube substratum becomes muddy, illustrate that bacterium grows, dull and stereotyped then dilution coating, picking has single bacterium of notable difference on colony characteristics, until obtaining single bacterial strain, and preserve in the slant medium ramp.
3, the screening of anti-chlorine ion dominant bacteria:
The above-mentioned isolated single bacterium colony of picking is inoculated in respectively in the screening culture medium, 35 ℃ of anaerobism constant temperature culture 5 days, and the muddy situation of observing bacteria suspension becomes the muddy high chlorine resistant ionic bacterial strain that filters out that is.Through screening, obtain the bacterial strain that a strain is numbered strain V 430 at last, i.e. anti-chlorine strain V 430.Anti-chlorine strain V 430 growing state on the liquid SC substratum that adds the different concns chlorion (is aided with [Cl simultaneously shown in Fig. 1-2
-OD during]=3000mg/L
600nmBe control value).
Two, the microorganism strains of anti-chlorine is identified:
1, to the evaluation of the stronger strain V 430 of chlorine-resistant property:
The stronger strain V 430 of chlorine-resistant property is carried out the evaluation of Physiology and biochemistry and the evaluation of 16S rDNA molecule, determined the kind of chlorine resisting strain from molecular level.
16S rDNA sequential analysis is mainly according to following steps:
1. the extraction of bacterium nuclear DNA,
1) choose single colony inoculation overnight incubation in LB liquid pipe with autoclaved toothpick, get 1.5ml bacterium liquid in the Eppendorf pipe, 8, the centrifugal 5min of 000rpm room temperature thoroughly removes supernatant.
2) add STE solution 1.5ml washing once, the centrifugal supernatant of abandoning adds 0.6mlTE solution again, resuspended bacterium.
3) add the N,O-Diacetylmuramidase of 30ul 10mg/ml, 37 ℃ of water-bath 45min,
4) add the SDS of 65 μ l 10%, the Proteinase K 3 μ l of 20mg/ml, 50 ℃ of water-baths 2 hours, clear to solution becomes.
5) add equivalance body phenol chloroform extracting three times, till can't see egg white layer.
6) NaAc (pH5.2) of the 3M of adding 1/10 volume in supernatant liquor.
7) add isopyknic Virahol, deposit D NA.Twine with glass stick and to choose the DNA filament, once with 70% washing with alcohol, natural airing, and DNA is dissolved in the 100 μ l TE solution.
8) adding final concentration is the RNase of 50 μ g/ml, and 37 ℃, 30min removes RNA, and-20 ℃ of preservations are standby.
2. the pcr amplification of 16S rDNA gene
P1: forward primer 5 ' AGAGTTTGATCCTGGCTCAG3 '
P2: reverse primer 5 ' GGTTACCTTGTTACGACTT3 '
In 50 μ L reaction volumes, add 1 μ L template DNA (0.1 μ g), 0.5 μ L P1 and P2 (final concentration is 0.5 μ M), 1 μ LdNTP (every kind of NTP0.2mM), 0.5 μ LTaq polysaccharase (2U) and 5 μ L, 10 * PCR damping fluid.The pcr amplification condition is: 94 ℃ of pre-sex change 5min; At 94 ℃ of sex change 30s, 61-65 ℃ of annealing 30s, 72 ℃ are extended 1min, circulate 30 times; Last 72 ℃ are extended 10min eventually.
3. the recovery of PCR product
After the PCR product carried out electrophoresis with 1% sepharose, under ultraviolet lamp, cut and contain desire and reclaim segmental gel, put into the 1.5ml centrifuge tube and add 2 times of volume TE, 65 ℃ of water-baths add the extracting of equal-volume water-saturated phenol and get the upper strata water after once centrifugal and use that phenol-chloroform-the primary isoamyl alcohol extracting once again after 10 minutes, reset and add 10mol/L ammonium acetate and 2 times of volume dehydrated alcohol precipitations of 0.1 times of volume in the collection, centrifugal, wash precipitation once with 70% ethanol, be dissolved in an amount of sterilization distilled water after air-dry.
4. the complete sequence determination of 16S rDNA and analysis
P-f：CACGACGTTGTAAAACGACAGTTTGATCCTGGCTC；
P-r：GGATAACAATTTCACACAGGAAGGAGGTGATCCAGCC。
Add 1 μ L template DNA (＜0.1 μ g), 30 circulations of pcr amplification (94 ℃ of 1min, 55 ℃ of 30s, 72 ℃ of 2min).Adopt dyestuff terminator termination reaction in the ABI PRISM sequencing kit.Then, on Applied Biosystem 373A dna sequencing instrument, check order.The 16S rDNA sequence that records adopts BLAST software and the comparative analysis of GenBank database, finally determines the kind of this bacterium from molecular level.
2,16S rDNA sequencing:
The present invention adopts the method for 16S rDNA sequencing and analysis is carried out the evaluation of molecular level to bacterium.Nuclear DNA with bacterium is a template, is primer with the universal primer of the pcr amplification of 16S rDNA gene, carries out pcr amplification, (detecting) with 1% agarose gel electrophoresis, and as Figure 1-3.After the PCR product is purified, measure its complete sequence.
＜110〉China Geological Univ. Wuhan
＜120〉gene of anti-chlorine strain V 430
<160>1423
<210>1
<211>1423bp
<212>DNA
＜213〉Staphylococcus saprophyticus (Staphylococcus saprophyticus)
<220>
<221>misc_feature
<222>
<400>1
GTTCCATGTGGGTAACCTACCTATAAGACTGTTCTAACTCCGGGATACCGGGGCTAATGC 120
CGGATGGCATTAGAACCGGTTGCCCGGAGTGTGAAAGATGGTTTTGCTATCTCTTATACG 180
TGGACCCGCGCCGTATGAGGTAGTTGGTAAGGTAATGGCTTACCAAGGCAACGATACGTA 240
GCCGACCTGAGAGGGTGATCGGCCACACTGGAACTGAGACACGGTCCAACTCTCTACGGG 300
AGGCAGCAGTAGGGAATCTTCCGCAATGGGCGAAAGCCTGACGGAGCAACGCCGCGTGAG 360
TGATGAAGGGTTTCGGCTCGTAAAACTCTGTTATTAGGGAAGAACAAACGTGTAAGTAAC 420
TGTGCACGTCTTGACGGTACCTAATCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGC 480
GGTAATACGTAGGTGGCAAGCGTTATCCGGAATTATTGGGCGTAAAGCGCGCGTAGGCGG 540
TTTCTTAAGTCTGATGTGAAAGCCCACGGCTCAACCGTGGAGGGTCATTGGAAACTGGGA 600
GACTTGAGTGCAGAAGAGGAAAGTGGAATTCCATGTGTAGCGGTGAAATGCGCAGAGATA 660
TGGAGGAACACCAGTGGCGAAAGCGACTTTCTGGTCTGTAACTGACGCTGATGTGCGAAA 720
GCGTGGGGATCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTA 780
AGTGTTAGGGGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGG 840
GAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGAG 900
CATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAAATCTTGACATCCTTTGACCA 960
CTCTAGAGATAGAGTTTTCCCCTTCGGGGGACAAAGTGACAGGTGGTGCATGGTTGTCGT 1020
CAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTAAACTTAGT 1080
TGCCAGCATTTAGTTGGGCACTCTAGGTTGACTGCCGGTGACAAACCGGAGGAAGGTGGG 1140
TACAAAGGGCAGCTAAACCGCGAGGTCATGCAAATCCCATAAAGTTGTTCTCAGTTCGGA 1260
TTGTAGTCTGCAACTCGACTACATGAAGCTGGAATCGCTAGTAATCGTAGATCAGCATGC 1320
TACGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAA 1380
CACCCGAAGCCGGTGGAGTAACCATTAATGGAGCTAGCCGTCG 1423
Three, the colonial morphology of anti-chlorine strain V 430, physiology and biochemical character see Table 1.Cellular form is seen Fig. 1-1a, Fig. 1-1b, Fig. 1-1c.
Table 1:
The colony morphology characteristic of strain V 430 and main physiological characteristic:
Nomenclature in the table: "+": the bacterial strain more than 90% is positive, "-": the bacterial strain more than 90% is negative
Four, strain V 430 is new bacterial strain:
The 16S rDNA gene order of strain V 430 and the sequence in the international GenBank database are carried out online homology find that relatively the homology of a plurality of bacterial strains of bacterium V430 and Staphylococcus saprophyticus (Staphylococcus saprophyticus) is up to more than the 98-99%.The Physiology and biochemistry and the Molecular Identification result of comprehensive strain V 430, can determine that bacterium V430 is Staphylococcus saprophyticus (Staphylococcus saprophyticus), strain V 430 called after Staphylococcus saprophyticus (Staphylococcussaprophyticus) V430 (being anti-chlorine strain V 430), consult pertinent data, do not have still that Staphylococcus saprophyticus belongs to organic wastewater with difficult degradation thereby under the relevant high-concentration chlorine ion condition and the report of its anti-chlorine capability study.Staphylococcus saprophyticus (Staphylococcussaprophyticus) V430 was new bacterial strain, had been preserved in Chinese typical culture collection center (being called for short CCTCC), preserving number: CCTCC NO.M205145 on December 9th, 2005.
The strain V 430 that the present invention separates, filters out has the function of better degrading high concentration chlorion difficult degradation saponin waste water.This has just widened people to the applied research thinking of Staphylococcus saprophyticus (Staphylococcus saprophyticus) in its function aspects, and provide useful bacterium source and technology for degraded contains the high-concentration chlorine ion saponin waste water, have stronger actual application value.
Five, the application of anti-chlorine strain V 430:
1. get saponin and produce composite waste, dilution 3-5 multiple, making its COD is 5000mg/L, uses CaCl
2Regulate [Cl
-] be 20,000 mg/L, get in the Erlenmeyer flask that waste water 50ml adds 250ml respectively, add the bacteria suspension 1ml of strain V 430,25-45 ℃ of constant temperature anaerobism cultivated, and the pH value is 7.5, reacts 3-5 days.Found that this bacterial strain can effectively remove the ability of COD in the saponin waste water, the COD that can remove in the waste water within 5d is about 50-60%.Factorial experiments sees Fig. 1-4a, Fig. 1-4b, Fig. 1-4c, Fig. 1-4d for details.
Described enlarged culturing base is: peptone 10g; Extractum carnis 5g; Distilled water 1000ml; CaCl
218.0g/L, pH=7.5.
2. the factorial experiments of Staphylococcus saprophyticus (Staphylococcus saprophyticus) V430 degraded saponin waste water:
1),, in waste water, adds CaCl to the removal effect of COD in order to investigate strain V 430 under different chlorine ion concentrations
29864.2,14980,21061,24780,30080mg/L the adjusting chlorine ion concentration is respectively:.Anti-chlorine strain V 430 removal effect result such as Fig. 1-4a.
From Fig. 1-4a as can be seen, as [Cl
-During]=9864.2mg/L, reaction times is 3d, strain V 430 only is 39.31% to the clearance of COD, but rising along with chlorine ion concentration in the waste water, strain V 430 increases the clearance of COD, when chlorine ion concentration in the waste water reaches 2.5 ten thousand mg/L, strain V 430 can reach 64.38% to the clearance of COD, and strain V 430 increases and changes little to the clearance of COD along with the continuation of chlorine ion concentration in the waste water, when chlorine ion concentration in the waste water during up to 30,000 mg/L, strain V 430 still remains on 60% to the clearance of COD.
2) in order to investigate bacterial strain (6610.4,7906.4,11209.6,13420.0,16196.2mg/L) under different COD concentration,, in waste water, add CaCl to the removal effect of COD
2Regulate chlorine ion concentration and be respectively 20000mg/L.Anti-chlorine strain V 430 removal effect result such as Fig. 1-4b.By experiment as can be known: [Cl in waste water
-]=20000mg/L, when COD concentration reaches 10,000 mg/L, strain V 430 is better to the clearance of COD, can reach 62.37%.
3) from Fig. 1-4c and 1-4d as can be seen, strain V 430 increases afterwards earlier with the increase of pH the clearance of COD and descends, and its highest clearance reaches 62% when pH is 7.5-8.Waste water after the centrifugal treating, when finding that pH is 7-8, the wet thallus amount is 9,10,11 o'clock apparently higher than pH, as seen during in neutrality, helps the growth of bacterial strain at pH; When inoculum size changed between 1-4ml, strain V 430 did not have considerable change to the clearance of COD, was 60% substantially, and when inoculum size was 1ml, its clearance was best.This is the certain nutritive substance of growth needs because of bacterial strain, and when bacteria suspension was 1ml, its material that needs was met easily.
＜110〉China Geological Univ. Wuhan
＜120〉anti-chlorine strain V 430 and screening method
<160>1423
<210>1
<211>1423bp
<212>DNA
＜213〉Staphylococcus saprophyticus (Staphylococcus saprophyticus)
<220>
<221>misc_feature
<222>
<400>1
GTTCCATGTGGGTAACCTACCTATAAGACTGTTCTAACTCCGGGATACCGGGGCTAATGC 120
CGGATGGCATTAGAACCGGTTGCCCGGAGTGTGAAAGATGGTTTTGCTATCTCTTATACG 180
TGGACCCGCGCCGTATGAGGTAGTTGGTAAGGTAATGGCTTACCAAGGCAACGATACGTA 240
GCCGACCTGAGAGGGTGATCGGCCACACTGGAACTGAGACACGGTCCAACTCTCTACGGG 300
AGGCAGCAGTAGGGAATCTTCCGCAATGGGCGAAAGCCTGACGGAGCAACGCCGCGTGAG 360
TGATGAAGGGTTTCGGCTCGTAAAACTCTGTTATTAGGGAAGAACAAACGTGTAAGTAAC 420
TGTGCACGTCTTGACGGTACCTAATCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGC 480
GGTAATACGTAGGTGGCAAGCGTTATCCGGAATTATTGGGCGTAAAGCGCGCGTAGGCGG 540
TTTCTTAAGTCTGATGTGAAAGCCCACGGCTCAACCGTGGAGGGTCATTGGAAACTGGGA 600
GACTTGAGTGCAGAAGAGGAAAGTGGAATTCCATGTGTAGCGGTGAAATGCGCAGAGATA 660
TGGAGGAACACCAGTGGCGAAAGCGACTTTCTGGTCTGTAACTGACGCTGATGTGCGAAA 720
GCGTGGGGATCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTA 780
AGTGTTAGGGGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGG 840
GAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGAG 900
CATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAAATCTTGACATCCTTTGACCA 960
CTCTAGAGATAGAGTTTTCCCCTTCGGGGGACAAAGTGACAGGTGGTGCATGGTTGTCGT 1020
CAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTAAACTTAGT 1080
TGCCAGCATTTAGTTGGGCACTCTAGGTTGACTGCCGGTGACAAACCGGAGGAAGGTGGG 1140
TACAAAGGGCAGCTAAACCGCGAGGTCATGCAAATCCCATAAAGTTGTTCTCAGTTCGGA 1260
TTGTAGTCTGCAACTCGACTACATGAAGCTGGAATCGCTAGTAATCGTAGATCAGCATGC 1320
TACGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAA 1380
CACCCGAAGCCGGTGGAGTAACCATTAATGGAGCTAGCCGTCG 1423
Claims (2)
1. a chlorine-resisting strain V430 is characterized in that described bacterial strain is Staphylococcus saprophyticus (Staphylococcussaprophyticus) V430 CCTCC NO.M205145.
2. a chlorine-resisting strain V430 according to claim 1, it is characterized in that: the colonial morphology of described Staphylococcus saprophyticus (Staphylococcus saprophyticus) V430 CCTCC NO.M205145: bacterium colony is rounded, children's oyster white in age, aged safran is opaque; Bacterium colony projection, edge-smoothing; Cell spheroid, d=0.6～single or paired, the in heaps appearance of 0.9 μ m, atrichia; Do not move no endogenous spore, Gram-positive, anaerobism; The suitableeest growth pH value: 6.0～7.5, growth temperature: 25-45 ℃; Main biochemical character: the catalase feminine gender, oxidase negative, glucose produce not aerogenesis of acid; Unhydrolysed casein, gelatin and starch; Can degrade fructose, seminose; Can reductive NO
3 -
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| CNB2006100181337A CN100422312C (en) | 2006-01-10 | 2006-01-10 | Chlorine resistant strain V430 and its screening method |
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Non-Patent Citations (6)
| Title |
|---|
| 3株油脂化工废水降解菌的分离鉴定. 黄钧等.应用与环境生物学报，第5卷第Suppl期. 1999 |
| 3株油脂化工废水降解菌的分离鉴定. 黄钧等.应用与环境生物学报，第5卷第Suppl期. 1999 * |
| 一株耐盐耐碱J101球菌的抗性研究. 刘爱民.安徽师范大学学报（自然科学版），第28卷第2期. 2005 |
| 一株耐盐耐碱J101球菌的抗性研究. 刘爱民.安徽师范大学学报（自然科学版），第28卷第2期. 2005 * |
| 降解邻硝基酚葡萄球菌的分离与特性研究. 卢涛等.四川大学学报（自然科学版），第37卷第Sup期. 2000 |
| 降解邻硝基酚葡萄球菌的分离与特性研究. 卢涛等.四川大学学报（自然科学版），第37卷第Sup期. 2000 * |
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