CN101864379A

CN101864379A - Radiation resistant pantoea sp. W36-1 and application thereof in environmental engineering

Info

Publication number: CN101864379A
Application number: CN 201010166704
Authority: CN
Inventors: 顾美英; 茆军; 谢玉清; 张志东
Original assignee: INSTITUTE OF MICROBIOLOGY XINJIANG ACADEMY OF AGRICULTURAL SCIENCES
Current assignee: INSTITUTE OF MICROBIOLOGY XINJIANG ACADEMY OF AGRICULTURAL SCIENCES
Priority date: 2010-05-10
Filing date: 2010-05-10
Publication date: 2010-10-20
Anticipated expiration: 2030-05-10
Also published as: CN101864379B

Abstract

The invention discloses pantoea sp. W36-1, of which the preservation number is CGMCC No. is 3701, a flocculant prepared by the pantoea sp. W36-1 and application of the flocculant in removal of suspensions, dyeing agents and heavy metal in waste water. Extracellular products of the bacteria have an obvious absorption peak at 200 to 250nm, and polysaccharide solution can remove 54.89 percent of free groups in the bacteria. The method for preparing the pantoea sp. W36-1 comprises the following steps of: performing fermentation culture of the bacteria strain for 48 hours, centrifuging 100ml of fermentation solution at a rotation speed of 6,000r/min for 10 minutes, adding absolute ethanol, of which the volume is 1.5 times that of supernatant, into the supernatant, uniformly mixing the absolute ethanol and the supernatant standing the mixed solution at the temperature of 4 DEG C overnight, centrifuging the precipitate, washing the precipitate by 70-percent ethanol for 2 to 3 times, and then drying the precipitate to obtain the finished product. The flocculent activity of the fermentation culture product of the pantoea sp. W36-1 is 81.7 to 83.15 percent, the yield of the pantoea sp. W36-1 is 6g/L and can be widely used for removing suspensions, dyeing agents, heavy metal and the like in waste water.

Description

General bacterium of a kind of radiation hardness and the application in environmental engineering thereof

Technical field

The invention belongs to the technical field of microbial strains resource applied environment engineering.Specifically, the technical field that relates to the application of a kind of general Pseudomonas novel species and applied environment engineering thereof.

Background technology

At present, in the technology and method of sewage disposal, flocculation sedimentation be widely used, one of lower-cost method, and the key in this process is to select and add the flocculation agent of excellent property for use.

Flocculation agent mainly contains three classes.The one, low molecule inorganic salts, as aluminium salt and molysite etc., large usage quantity easily causes secondary pollution, influences HUMAN HEALTH; The 2nd, synthetic polymeric flocculant, flocculation rate is fast, efficient height, but have the biological degradation difficulty, problem such as residual monomer is poisonous; The 3rd, natural organic high-molecular flocculant has that selectivity is strong, nontoxic, low price, be easy to advantages such as degraded, non-secondary pollution and regeneration, is the focus of present flocculation agent research, further applies but more weak flocculation activity has limited it.

Natural organic high-molecular flocculant mainly comprises compositions such as glycoprotein, mucopolysaccharide, Mierocrystalline cellulose and ribose by microorganisms, and flocculation agent composition that each strain bacterium is produced and characteristic be difference to some extent all.At present, the microorganism of the produced flocculation agent of having reported has bacterium [as rhodococcus (Rhodococcus), coryneform bacteria (Corynebacterium), pseudomonas (Pseudomonas), bacillus (Bacillus), Agrobacterium (Agrobacterium) etc.], actinomycetes [as Nocardia bacteria (Nocardia), streptomycete (Streptomyces)], fungi [as debaryomyces hansenii (Hansenula), fission yeast (Schizosaccharomyces), stalk mould (Aureobasidium), mould, aspergillus etc.] three major types.

The flocculation agent of microorganisms generally has the flocculating settling effect to the particulate state suspended substance of aqueous phase, the flocculation agent extract that produces as isolated bacterial M-127 in the active sludge can improve sewage at dehydrating effect (Zhao Jihong, Yang Jingfeng etc. microbial flocculant improves the research of dewatering performance of sludge. environmental science and technology, 2009,11:88-90.).The flocculation agent of microorganisms has the characteristic of absorption staining agent and heavy metal ion, as microbial flocculant called after mbf1 soluble pigment methyne orchid had decolorizing effect (Wu Juan preferably, Fei Wenyan. the flocculating properties of microbial flocculant and the research of decoloring ability thereof. the biology magazine, 2008,2:30-32,10.), the microbial flocculant (MBF) that colloid bacillus cereus produces has different throwing out (Yao Minjie to the different metal ion in the high density heavy metal ion simulated wastewater, connect the guest, microbial flocculant is studied high density effluent containing heavy metal ions throwing out. environmental science and technology, 2009,11:1-4.).

General Pseudomonas (Pantoea) is under the jurisdiction of bacterium microbe, has multiple biological characteristics and function, is the quasi-microorganism with great commercial exploitation prospect.Found in fields such as medicine, makeup research and development its can produce the anti-ultraviolet radiation material [Wang Hongyuan etc. anti-uv b radiation bacterium uv-absorbing meta-bolites is analyzed and the anti-ultraviolet radiation activity research. Chinese Journal of Marine Drugs, 2006,25 (4): 1-5], can produce immunomodulator [calm etc. the separation and purification and the evaluation of the low molecule lipopolysaccharides of pantoea agglomerans. Guangxi Medical University's journal, 2003,20 (6): 840-842]; Field of environment engineering find the degradable malachite green bacterial strain [Ren Qian etc. the isolation identification of malachite green degradation bacteria M3 and degradation characteristic research. ecological and rural environment journal, 2007,23 (3): 65-69], and the bacterial strain M-127[Yang Jing peak etc. that can produce flocculation agent. the optimization of pantoea agglomerans producing microbial flocculation agent flocculating conditions. the Speciality Petrochemicals progress, 2009,10 (2): 39-42].Along with research go deep into new bacterial classification and characteristic just progressively is studied and uses.

Summary of the invention

The object of the present invention is to provide the new bacterium of the general Pseudomonas of a kind of new Microbial resources, and its potential commercial exploitation prospect is described from field of environment engineering technology.

The present invention specifically provides the general bacterium W36-1 of strain separation screening from the desert Environment of Xinjiang, and this bacterium has stronger ultraviolet light resistant characteristic, detects through microbial taxonomy and identifies, is defined as general Pseudomonas novel species.At present, this bacterial strain was preserved in the international depositary institution of budapest treaty microorganism before the applying date: China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC).The address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode: 100101, preservation date is on March 26th, 2010, preserving number is CGMCC No.3701.Determine that it is general Pseudomonas novel species through the evaluation of microorganism polyphase sort, its biology title is tentative to be general bacterium (Pantoea sp.W36-1), determines the classification called after Pantoea sp. of this bacterium suggestion.

General bacterium (Pantoea sp.W36-1) CGMCC No.3701 strain growth is on solid PDA substratum, take by weighing the 200g potato, clean peeling is cut into small pieces, add water 1000ml and boil half hour, filtered through gauze adds 20g glucose, sterilizes about 20 minutes for 121 ℃, rounded, smooth moistening, protruding, the pale pink in bacterium colony surface, the sticky picking that is difficult for are transparent; Gram-negative (G-), aerobic, cell is shaft-like, do not form gemma; Can in 15-45 ℃ of mesophilic range, grow.

16S rRNA gene to the W36-1 bacterial strain checks order and compare of analysis, and itself and general Pseudomonas (Pantoea) homology similarity are the highest, with Pantoea gaviniae A18/07T highest homology be 97.1%.

According to " common bacteria identification handbook ", " microorganism classification such as uncle's Jie Shi Bacteria Identification handbook is identified the result that criterion and polyphase sort are identified, determine that the W36-1 bacterial strain is general Pseudomonas one novel species of bacterium class, the general bacterium of called after (Pantoea sp.W36-1) is determined the classification called after Pantoea sp. that this bacterium is advised.

Bacterial strain W36-1 thalli growth provided by the invention does not have particular requirement with breeding to nutritive substance, all can grow on substratum such as common bacteria culture medium and PDA, Cha Shi.Conventional microbial preservation technology and methods such as method, freeze-drying, glycerine pipe carry out preservation and viability detects can to adopt the inclined-plane to go down to posterity.

With the W36-1 inoculation in the Pb that contains different concns ²⁺, Cu ²⁺, Co ²⁺, Hg ²⁺, Zn ²⁺PDA solid medium flat board on, 30 ℃ of constant temperature culture are observed, the W36-1 bacterial strain is to Co ²⁺Tolerance lower, and higher to other metal ion tolerance of measuring; To removing Co ²⁺Four heavy metal species ions in addition all have certain adsorption, wherein to Pb ²⁺The ionic adsorption effect is best, and adsorption rate reaches 89.2%; Hg ²⁺Secondly, be 21.76%, Cu ²⁺Be 12.45%; The W36-1 extracellular products has an obvious absorption peaks at 200-250nm, and polysaccharide soln can be eliminated its free radical of 54.89%.

Further, the invention provides general bacterium (Pantoea sp.W36-1) CGMCC No.3701 and prepare the preparation method of flocculation agent, after the general bacterium of bacterial strain (Pantoea sp.W36-1) CGMCC No.3701 carried out fermentation culture 48h, get 100ml fermented liquid centrifugal 10min under the 6000r/min condition, supernatant liquor adds the dehydrated alcohol of 1.5 times of volumes, 4 ℃ of following standing over night behind the mixing, the throw out of separating out is centrifugal 30min under the 6000r/min condition, the throw out of gained is with 70% washing with alcohol 2～3 times, and vacuum-drying or constant temperature air seasoning get the flocculation agent crude product.

General bacterium (Pantoea sp.W36-1) CGMCC No.3701 fermenting culture has flocculation activity preferably, and the fermenting culture flocculation activity is 81.7%-83.15%, and its productive rate is 6g/L.Institute's produce flocculant all has decolorizing effect preferably to magenta, haematochrome (self-control microorganism natural red colouring matter), staining agents such as brooethyl phenol is green, chrome black is blue, methylene blue, especially natural red colouring matter, brooethyl phenol is green, chrome black is blue percent of decolourization all is higher than 90%.Illustrate that the flocculation agent that the W36-1 bacterial strain is produced can use aspect some staining agent wastewater treatment.

By implementing the concrete summary of the invention of the present invention, can reach following beneficial effect:

Directed screening of the present invention is to the flocculation agent of general bacterium (Pantoea sp.W36-1) CGMCC No.3701 and generation thereof and the application aspect suspended substance, staining agent and the heavy metal of flocculation agent in removing waste water.Show that after testing the fermenting culture flocculation activity reaches as high as 81.7%-83.15%, its productive rate is 6g/L; Natural red colouring matter, brooethyl phenol is green, chrome black is blue percent of decolourization all are higher than 90%; Remove Co ²⁺Pb in addition ²⁺, Hg ²⁺, Cu ²⁺, Zn ²⁺Four heavy metal species ions all have certain adsorption, wherein to Pb ²⁺The ionic adsorption effect is best, and adsorption rate reaches 89.2%.

Description of drawings:

Figure 1 shows that general bacterium (Pantoea sp.W36-1) CGMCC No.3701 uvioresistant characteristic.

Figure 2 shows that the adsorptive power of general bacterium (Pantoea sp.W36-1) CGMCC No.3701 culture to different heavy metals.

Figure 3 shows that the decolorizing effect of general bacterium (Pantoea sp.W36-1) CGMCC No.3701 culture to different pigments.

Figure 4 shows that general bacterium (Pantoea sp.W36-1) CGMCC No.3701 growth and fermenting culture flocculation activity.

Figure 5 shows that general bacterium (Pantoea sp.W36-1) CGMCC No.3701 extract ultraviolet-visible spectrophotometric scintigram.

Embodiment

Below be the application experiment of bacterial strain provided by the invention, be intended to illustrate the one side of its application prospect, and the Application Areas of bacterial strain provided by the present invention, application method are not limited thereto in field of environment engineering technology.

Invention is adopted following basic medium if no special instructions in following examples.

The LB substratum: peptone 10g, yeast soak powder 5g, NaCl 5g, water 1000ml, pH7.0.

The PDA substratum: take by weighing the 200g potato, clean peeling is cut into small pieces, and adds water 1000ml and boils half hour, and filtered through gauze adds 20g glucose, sterilizes about 20 minutes for 121 ℃.

Czapek's solution: NaNO ₃3g, K ₂HPO ₄1g, MgSO ₄7H ₂O 0.5g, KCl 0.5g, FeSO ₄7H ₂O 0.01g, sucrose 30g, H ₂O 1000ml, heating for dissolving, 121 ℃ of sterilization 20min after the packing.

Embodiment 1: cultivation, the evaluation of general bacterium (Pantoea sp.W36-1) CGMCC No.3701

By the general bacterium W36-1 of separation screening from the desert Environment of Xinjiang, this strain growth is in solid PDA substratum, by taking by weighing the 200g potato, clean peeling is cut into small pieces, and adds water 1000ml and boils half hour, filtered through gauze, add 20g glucose, sterilized 20 minutes for 121 ℃.The W36-1 bacterial strain be Gram-negative (G-), aerobic, cell is shaft-like, do not form gemma; Can well grow up on LB, PDA and czapek's solution, strain growth is at solid PDA substratum, and rounded, smooth moistening, protruding, the pale pink in bacterium colony surface, the sticky picking that is difficult for are transparent; Gram-negative (G-), aerobic, cell is shaft-like, do not form gemma; Can in 15-45 ℃, grow.Has stronger ultraviolet light resistant characteristic.Determine that it is general Pseudomonas novel species through the evaluation of microorganism polyphase sort, its biology title is tentative to be Pantoea sp.W36-1.This bacterial strain has been preserved in the international depositary institution of budapest treaty microorganism at present: China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC).The address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode: 100101, preservation date is on March 26th, 2010, preserving number is CGMCC No.3701.

The oxydase reaction feminine gender; With BIOLOG MICROSTSTION SYSTEM GN2 test card its metabolic characteristics is carried out determination and analysis, the result is as shown in table 1.

According to " common bacteria identification handbook ", " microorganism classification such as uncle's Jie Shi Bacteria Identification handbook is identified and the result that criterion and polyphase sort are identified is determined that the W36-1 bacterial strain is general Pseudomonas one novel species of bacterium class.

Table 1 Biolog Microstation System GN2 test card test result

Embodiment 2: the anti-ultraviolet radiation characteristic of general bacterium (Pantoea sp.W36-1) CGMCC No.3701

Is basic medium with the W36-1 bacterial strain with liquid LB, 30 ℃, 180r/min constant temperature vibration training oxygen 16h, to the bacterial classification logarithmic phase after.4000rpm collects the thalline in the culture, is OD to suspend with physiological saline and to dilute behind twice of physiological saline (0.75%NaCL) the solution washing thalline ₆₀₀It is 0.2 bacteria suspension; Get the 10mL bacteria suspension and place the 9cm culture dish, be positioned over 25cm place under the 15W ultraviolet lamp, the vibration radiation treatment is taken a sample 1 time at set intervals, detects the survival volume of bacterial strain with spread plate, and is contrast with intestinal bacteria, and the result is referring to shown in the accompanying drawing 1.

Measurement result shows that W36-1 still has survival behind the uv-radiation 15min under these conditions, and corresponding intestinal bacteria do not see survival behind radiation 1min, this shows that the W36-1 bacterial strain is had stronger anti-ultraviolet radiation characteristic.

Embodiment 3: general bacterium (Pantoea sp.W36-1) CGMCC No.3701 counterweight metal ion tolerance

The W36-1 bacterial strain is inoculated in the Pb that contains different concns respectively ²⁺, Cu ²⁺, Co ²⁺, Hg ²⁺, Zn ²⁺PDA solid medium flat board on, 30 ℃ of constant temperature culture detect its upgrowth situation.Its measurement result is as shown in table 2.

Table 2W36-1 bacterial strain is to the tolerance experiment (concentration unit mg/L) of different heavy metals

	??50	??100	??200	??300	??400	??500	??600	??700	??800	??900	??1000	??1100	??1200
	??50	??100	??200	??300	??400	??500	??600	??700	??800	??900	??1000	??1100	??1200	??Pb ²⁺	??+	??+	??+	??+	??+	??+	??+	??+	??+	??+	??+	??+	??+
??Cu ²⁺	??+	??+	??+	??+	??±	??-	??-	??-	??-	???-	??-	-	?-	??Pb ²⁺	??+	??+	??+	??+	??+	??+	??+	??+	??+	??+	??+	??+	??+
??Cu ²⁺	??+	??+	??+	??+	??±	??-	??-	??-	??-	???-	??-	-	?-	??Co ²⁺	??-	??-	??-	??-	??-	??-	??-	??-	??-	???-	??-	-	?-
??Hg ²⁺	??-	??+	??-	??+	??-	??±	??-	??-	??-	???-	??-	-	?-	??Co ²⁺	??-	??-	??-	??-	??-	??-	??-	??-	??-	???-	??-	-	?-
??Hg ²⁺	??-	??+	??-	??+	??-	??±	??-	??-	??-	???-	??-	-	?-	??Zn ²⁺	??+	??+	??+	??+	??+	??+	??+	??+	??+	??±	??±	-	?-

Annotate :+be growth ,-for not growing, ± for a little less than growing

By table 2 result as can be known, under experiment condition the W36-1 bacterial strain to Pb ²⁺Tolerance more than 1200mg/L, Cu ²⁺At 400mg/L, Hg ²⁺At 500mg/L, Zn ²⁺At 1000mg/L, Co ²⁺Tolerance less than 50mg/L.Illustrate that the W36-1 bacterial strain is to Co ²⁺Tolerance lower, and higher to other metal ion tolerance of measuring.

Embodiment 4: general bacterium (Pantoea sp.W36-1) CGMCC No.3701 is to the absorption of heavy metal ion

In bacterial strain tolerance scope, preparation different concns Pb ²⁺, Cu ²⁺, Co ²⁺, Hg ²⁺, Zn ²⁺The PDA liquid fermentation medium, its concentration is respectively 1000mg/L, 300mg/L, 800mg/L, 50mg/L and 700mg/L, insert the W36-1 bacterial strain in 30 ℃, 180r/min constant-temperature shaking culture 48 hours, the centrifugal 5min of 6000r/min, get supernatant liquor and measure the content of its different heavy metals, calculate the adsorptive power of this bacterium each heavy metal.Its adsorption rate calculation formula is: r=(B ₀-B ₁)/B ₀* 100%, in this formula, B ₀Be the concentration of heavy metal in the substratum that does not add bacterium liquid, B ₁For adding the concentration of the heavy metal after bacterium liquid is cultivated.The result is referring to shown in the accompanying drawing 2.

The measuring method of different heavy metal ion carries out with reference to the method for " analytical applications of EDTA ex hoc genus anne compound " (the high minister of Zhao is translated, Geology Publishing House, 1982 for Pu Xi ratio, R. work).Pb ²⁺, Cu ²⁺, Co ²⁺, Zn ²⁺Adopt the xylenol orange development process, Hg ²⁺Adopt methyl thymol blue development process.

By accompanying drawing 2 results as can be known the W36-1 bacterial strain to removing Co ²⁺Four heavy metal species ions in addition all have certain adsorption, wherein to Pb ²⁺The ionic adsorption effect is best, and adsorption rate reaches 89.2%; Hg ²⁺Secondly, be 21.76%, Cu ²⁺Be 12.45%.

Embodiment 5: general bacterium (Pantoea sp.W36-1) CGMCC No.3701 prepares the adsorption of flocculation agent to staining agent

In the PDA liquid fermentation medium, in 30 ℃, 180r/min constant-temperature shaking culture 48 hours, directly the absorption property that carries out staining agent with fermenting culture was measured with the W36-1 inoculation.Concrete measuring method: at the magenta of 10mL, 0.05g/l, haematochrome (self-control microorganism natural red colouring matter), brooethyl phenol is green, chrome black is blue, respectively add 10mL fermenting culture, 2mL 1% CaCl in the methylene blue solution (be with pH 7 buffer preparation) ₂Solution leaves standstill more than the 10min behind the vibration mixing; Respectively at λ _540nm, λ _530nm, λ _620nm, λ _635nm, λ _620nmThe place measures its light absorption value, and is blank with the pigment solution that does not add fermenting culture, calculates percent of decolourization r=(A ₀-A ₁)/A ₀* 100%, in this formula, A ₀For not adding the absorbance of bacterium liquid, A ₁For adding the absorbance of bacterium liquid.The result is referring to shown in the accompanying drawing 3.

Show that referring to measurement result shown in the accompanying drawing 3 W36-1 institute produce flocculant all has decolorizing effect preferably to above-mentioned several pigments, especially natural red colouring matter, brooethyl phenol is green, chrome black is blue percent of decolourization all are higher than 90%.Illustrate that the flocculation agent that the W36-1 bacterial strain is produced can use aspect some staining agent wastewater treatment.

Embodiment 6: general bacterium (Pantoea sp.W36-1) CGMCC No.3701 fermenting culture flocculation activity

In the PDA liquid fermentation medium, in 30 ℃, 180r/min constant-temperature shaking culture, every 6h sampling is not done the reference zeroing to inoculate the culture of strains base, measures nutrient solution at OD with the W36-1 inoculation ₆₀₀Growth curve, measure nutrient solution simultaneously in flocculation activity to the kaolin suspension liquid.

Flocculation activity detection method: get fermenting culture 2mL, add among the kaolin solution 50mL of 1g/L, leave standstill and handle more than 5 minutes, with λ _550nmLight wave detects its absorbancy, and its light absorption value note is made A; With blank fermention medium is blank, and its light absorption value note is made B; Flocculation clearance (F) is: [(B-A) ÷ B * 100%].Measurement result is referring to shown in the accompanying drawing 4.

Show that by the growth curve shown in the accompanying drawing 4 W36-1 enters logarithmic phase in 12h, enter stationary phase behind the 24h and rise slowly that its OD550 slightly reduces behind the 42h; The flocculation curve shows that the flocculating rate of W36-1 rises gradually along with the growth of bacterium before 24h, but 24-42h slightly descends, but in the 48h fast rise, flocculating rate reaches maximum 80.28%, descends fast subsequently.

Embodiment 7: general bacterium (Pantoea sp.W36-1) CGMCC No.3701 prepares the extraction of flocculation agent and the flocculation activity of extract

Press the method shown in the embodiment 6, after bacterial strain W36-1 carried out fermentation culture 48h, get 100ml fermented liquid centrifugal 10min under the 6000r/min condition, supernatant liquor adds the dehydrated alcohol of 1.5 times of volumes, 4 ℃ of following standing over night behind the mixing, the throw out of separating out is centrifugal 30min under the 6000r/min condition, and the throw out of gained is with 70% washing with alcohol 2～3 times, and vacuum-drying or constant temperature air seasoning get the flocculation agent crude product.Weigh and calculate the flocculation agent productive rate; Prepared flocculation agent is dissolved in the 100ml water again, is mixed with the aqueous solution, carry out the detection of flocculation activity, detect the fermenting culture flocculation activity simultaneously, difference more between the two by method described in the embodiment 6.

After measured, carry out the production and the extraction of flocculation agent with the method described in the present embodiment, the fermenting culture flocculation activity is 81.7%, and its productive rate is 6g/L, and Zhi Bei aqueous solution flocculation activity brings up to 83.15% again.

The measuring presentation of results prepares flocculation agent with the described method of present embodiment, can extract the effective constituent that fermentation culture produces; Leaching process may be removed partial impurities and help to improve flocculation activity; Fermenting culture has identical flocculation activity with extract, can directly apply to water and handle.

Embodiment 8: different substratum are to flocculation agent and active influence thereof

With the czapek's solution is fermention medium, carries out the preparation and the active comparative analysis of fermentation culture, flocculation agent by the method described in the embodiment 6,7.

This measuring result shows, carry out fermentation culture with czapek's solution, its fermenting culture flocculation activity can reach 82.78%, but it prepares the flocculation agent productive rate by method described in the embodiment 7 0.3g/L is only arranged, and the aqueous solution flocculation activity of preparation again only has 35.7%.The flocculation agent that two kinds of substratum fermentation culture of this result of implementation and 6,7 detected result comparative descriptions of enforcement are produced has bigger otherness on main composition; Can produce flocculation agent of different nature by the control of substratum nutrition source.

Embodiment 9: the photoabsorption and the anti-oxidation characteristics of general bacterium (Pantoea sp.W36-1) CGMCC No.3701 extracellular products

Carry out the fermentation culture of bacterial strain and the extraction of meta-bolites by the method described in the embodiment 6,7.The product that extracted with water dissolution, is carried out all wave band (or carrying out the photoscanning analysis under the ultraviolet band) under uv-spectrophotometric, and its measurement result is referring to shown in the accompanying drawing 5;

Carry out the mensuration of anti-hydroxy radical qiao oxidation characteristic with the Fenton method.The 0.15mo l/LFeSO that adds 1ml ₄Solution, the 2mmol/L Whitfield's ointment of 0.3ml, 1ml 100mg/L polysaccharide solution adds 0.7ml 6mmol/L H at last ₂O ₂Start, 25 ℃ of reaction 1h measure OD510nm, and by formula free radical scavenging activity (%)=[1-(Ai-A0i)/A0] * 100% calculates, and wherein A0 does not add sample for contrast, and Ai is the light absorption value of sample, and A0i is not for adding the light absorption value of Whitfield's ointment sample.

By accompanying drawing 5 as can be seen, extracellular products of the present invention has an obvious absorption peaks at 200-250nm.Fenton method measurement result shows that this polysaccharide soln can eliminate 54.89% free radical.

Pass through the foregoing description, can prove absolutely general bacterium (Pantoeasp.W36-1) the CGMCC No.3701 of directed screening provided by the invention, the carbon nitrogen source of available routine carries out fermentation culture, shows that after testing the fermenting culture flocculation activity reaches as high as 81.7%-83.15%, and its productive rate is 6g/L; Natural red colouring matter, brooethyl phenol is green, chrome black is blue percent of decolourization all are higher than 90%; Remove Co ²⁺Pb in addition ²⁺, Hg ²⁺, Cu ²⁺, Zn ²⁺Four heavy metal species ions all have certain adsorption, wherein to Pb ²⁺The ionic adsorption effect is best, and adsorption rate reaches 89.2%.Flocculation agent can be widely used in the suspended substance and the staining agent field in removing waste water of removing in the waste water, has widely aspect the heavy metal of Zhi Bei flocculation agent in removing waste water simultaneously and uses.

Claims

1. a general bacterium (Pantoea sp.W36-1) CGMCC No.3701.

2. general bacterium as claimed in claim 1 (Pantoea sp.W36-1) CGMCC No.3701 is characterized in that described bacterial strain is to Co ²⁺Tolerance lower, and higher to other metal ion tolerance.

3. general bacterium as claimed in claim 1 (Pantoea sp.W36-1) CGMCC No.3701 is characterized in that described bacterial strain is to Pb ²⁺Adsorption rate is 89.2%; Hg ²⁺Adsorption rate is 21.76%, Cu ²⁺Adsorption rate is 12.45%.

4. general bacterium as claimed in claim 1 (Pantoea sp.W36-1) CGMCC No.3701 is characterized in that described bacterial strain extracellular products is at the 200-250nm absorption peak, and polysaccharide soln has the elimination free radical.

5. general bacterium as claimed in claim 1 (Pantoea sp.W36-1) CGMCC No.3701 produces the preparation method of flocculation agent, it is characterized in that, described flocculation agent is the flocculation agent crude product, after the general bacterium of bacterial strain (Pantoeasp.W36-1) CGMCC No.3701 carried out fermentation culture 48h, get 100ml fermented liquid centrifugal 10min under the 6000r/min condition, supernatant liquor adds the dehydrated alcohol of 1.5 times of volumes, 4 ℃ of following standing over night behind the mixing, the throw out of separating out is centrifugal 30min under the 6000r/min condition, the throw out of gained is with 70% washing with alcohol 2～3 times, and vacuum-drying or constant temperature air seasoning get the flocculation agent crude product.

6. the flocculation agent that general bacterium as claimed in claim 1 or 2 (Pantoea sp.W36-1) CGMCC No.3701 produces.

7. the application of the flocculation agent that general bacterium as claimed in claim 6 (Pantoea sp.W36-1) CGMCC No.3701 produces in removing waste water suspension.

8. the application of the flocculation agent that general bacterium as claimed in claim 6 (Pantoea sp.W36-1) CGMCC No.3701 produces in removing the waste water staining agent.

9. the application of the flocculation agent that general bacterium as claimed in claim 6 (Pantoea sp.W36-1) CGMCC No.3701 produces in aspect removing the waste water heavy metal.