CN103103146B - The Sphingomonas of one strain degraded bromate and application thereof - Google Patents
The Sphingomonas of one strain degraded bromate and application thereof Download PDFInfo
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Abstract
The invention discloses Sphingomonas (Sphingomonas sp.815Y) bacterial strain and the application thereof of a strain degraded bromate, its microbial preservation number is CGMCC No.4721.Sphingomonas strain of the present invention obtains from occurring in nature screening, can effectively to degrade bromate, degradation efficiency is high, can inoculate and be applied in BAC process, bromate in the bromate generated in effective removal tap water ozonation and contaminated tap water, improve the security of tap water, have a good application prospect in drinking water treatment.
Description
Technical field
The present invention relates to the microorganism strains that noxious pollutant is removed in a strain, particularly relate to Sphingol single-cell (Sphingomonas sp.) bacterial strain of a strain degraded bromate, belong to the microbiological deterioration field of bromate.
Background technology
Along with the development of Water purification technology and people are to the attention of drinking-water quality, the application of ozonation technology in tap water is increasingly extensive.Ozone all has obvious inactivating efficacy to most bacterium, virus, fungi and protozoon, egg capsule, can remove the multiple traces of harmful pollutent such as disinfection byproduct (DBP) precursor, Taste and odor compounds in water efficiently simultaneously.But some by products produced in ozonation, certain negative impact is also caused to drinking water safety, as can bromate be produced in brominated former water-ozone process, there is carinogenicity, and to be difficult to remove by subsequent technique, limit the further application of ozone to a certain extent.
Bromate under field conditions (factors) little in colorless and odorless, volatility, be not hydrolyzed, the character such as thermally-stabilised, because it has the genetoxic of DNA and Chromosome level, bromate is regarded as the potential carcinogen of 2B level by international cancer research institution.The research of US Gov Env Protection Agency shows: the content of bromate in drinking water is that μ g/L level all can have certain carcinogenesis.The carcinogenic risk rate taking in the bromate of 3 μ g/L is 10
-5; The carcinogenic risk rate taking in the bromate of 5 μ g/L is 10
-4.In the world, in the tap water of the World Health Organization and EPA's defined, bromate maximum permissible concentration is within 0.01mg/L, and this index in Chinese drinking water standard is consistent with international standard.
For the bromate occurred in tap water ozonation, Chinese scholars have studied a lot of minimizing technology, such as ammonification, adds hydrogen peroxide, oxidation-reduction, conductor photocatalysis, novel metal catalyzer, ion-exchange membrane etc.Although aforesaid method can remove bromate to a certain extent, deposit problem in many applications, as high cost, cause secondary pollution etc.Membrane processing method is better to the removal effect of bromate, but cost is very high, and can produce some waste water containing the bromate of high density needs process in addition simultaneously; The removing of active carbon adsorption to bromate also has better effects, but need periodic replacement and costly, troublesome poeration; Adopt low pressure, although medium-pressure or high pressure mercury lamp UV-light appropriateness can decompose bromate, it has more also decomposed ozone reserved before bottling simultaneously.More to the research of removing bromate in water, the application for a patent for invention being CN101439900A as publication number discloses the method and system that bromate in water is removed in a kind of redox.The method is: first is in lower level by the dissolved oxygen concentration in the nitrification control water of microorganism on ammonium salt and gac; Second by the bromate in the reductibility removal water of ferrous sulfate; 3rd removes iron ion excessive in water by filtering unit.The system of this removal bromate comprises: control the pretreatment work section of oxygen in water, remove the filtration treatment working zone of the redox working zone of bromate and removal excessive iron ion, wherein, described pretreatment work section is connected with described redox working zone, and described redox working zone is connected with described filtration treatment working zone.Although the method can reach more than 60% to the clearance of bromate, this treatment process need control dissolved oxygen, is difficult to application in actual process, and complex operation, easily causes secondary pollution simultaneously.
Work in the biological degradation of bromate is in progress less both at home and abroad.For this reason, filter out bacterial strain bromate being had to efficient degradation ability from occurring in nature, the removal being applied to bromate in water will have actual using value.
Summary of the invention
The present invention is directed to prior art Problems existing, the present invention screens a strain Sphingol single-cell from occurring in nature, this Sphingomonas can effectively be degraded bromate, bromate in the tap water removing the bromate that occurs in tap water ozonation and polluted by bromate, has a good application prospect in drinking water treatment.
For achieving the above object, one aspect of the present invention provides a strain Sphingol single-cell Sphingomonas sp.815Y.
Sphingomonas provided by the present invention is preserved in " China Committee for Culture Collection of Microorganisms's common micro-organisms " center " on March 28th, 2011; Its microbial preservation number is CGMCC No.4721; Its Classification And Nomenclature is: Sphingomonas sp.815Y, and the preservation time is on March 28th, 2011; Preservation address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica.
Sphingomonas of the present invention adopts the separation of enrichment culture, plate dilution method from the charcoal of the BAC process section of Environmental Biotechnology study group of Ecological Environment Research Center, Chinese Academy of Sciences (environmental technology building, No. 18, Haidian District Beijing Shuan Qing road, postcode 100085) ozone biological activated carbon small testing device, purifying obtains.
Wherein, substratum composed as follows of enrichment culture: CaCl
25mg, MgCl
25mg, MnCl
20.3mg, FeCl
20.3mg, ZnCl
20.01mg, H
3bO
30.01mg, CoCl
20.005mg, bromate 1mg, sodium acetate 10mg, distilled water 1,000mL, pH 7.0-8.0, substratum constant temperature sterilizing 20 minutes at 121 DEG C.
Wherein, substratum composed as follows of isolation and purification culture: yeast extract 0.5g, Tryptones 0.5g, acid hydrolysis casein 0.5g, glucose 0.5g, Zulkovsky starch 0.5g, K
2hPO
40.3g, MgSO
47H
2o 0.05g, Sodium.alpha.-ketopropionate 0.3g, agar 15g, pH 7.0-8.0, substratum constant temperature sterilizing 20 minutes at 121 DEG C.
Sphingomonas Sphingomonas sp.815Y of the present invention has following characteristics:
1, Sphingomonas Sphingomonas sp.815Y is suitable for growing in the developing medium of neutral meta-alkalescence (pH7-8), the growth temperature be applicable to is 15-35 DEG C, its bacteriology morphological specificity is: cell is shaft-like, and size is 0.53 ~ 0.70 μm × 1.06 ~ 1.67 μm; Gram-negative; Not sprout spore, atrichia.
2, Sphingomonas colonial morphology on LB with nutrient agar flat board is consistent, cultivates the bacterium colony of 2 days in yellow, moistening, and droplet-like, circular, protruding, opaque, edge is comparatively neat.
3,16S rRNA gene sequence characteristic is: the 16S rRNA sequence length of bacterial strain Sphingol single-cell Sphingomonas sp.815Y is 1466bp.
Wherein, strain screening in accordance with the following steps:
With the biological activated carbon of the ozone-BAC process section of the small testing device of Environmental Biotechnology study group of Ecological Environment Research Center, Chinese Academy of Sciences processing drinking water for sample, carry out enrichment culture as bacterium source.Described small testing device is used for advanced treatment tap water, comprise the ozonation technology section and bioactive carbon absorbing process section two portions of carrying out successively, ozonation technology wherein utilizes ozone as advanced oxidation agent, by the hard-degraded substance degraded cannot removed in traditional drinking water treatment technique, larger molecular organics is decomposed into small organic molecule; Bioactive carbon absorbing process to adsorb small-molecule substance with gac and microorganism and decomposes, thus reach the object of purifying drinking water.When gac just brings into operation, carbon surface is not biological, and along with the prolongation of working time, the microbes in water and in air is enriched in formation on bioactive carbon gradually and has certain thickness microbial film, becomes stable bioactive carbon.Collection of the present invention is biological activated carbon is gather after the bioactive carbon absorbing process section of small testing device runs 5 months continuously.
Gathered biological activated carbon sample is mixed with phosphate buffered saline buffer, makes bacteria suspension; Then, the centrifugate after bacteria suspension is centrifugal is inoculated in enrichment medium carries out enrichment culture; Then enrichment culture thing is coated on plate isolation pure medium and repeatedly carry out flat board coating, carry out isolation and purification culture, obtain Sphingomonas strain bromate to removal ability until be separated.
Wherein, described phosphate buffered saline buffer is the mixing solutions of Sodium phosphate dibasic and SODIUM PHOSPHATE, MONOBASIC, and pH is 7.2.
Particularly, the compound method of described phosphate buffered saline buffer is: in 800ml distilled water, add 8gNaCl, 0.2g KCl, 1.44g Sodium phosphate dibasic and 0.24g potassium primary phosphate, with salt acid for adjusting pH to 7.2, add water and be settled to 1L, at 121 DEG C constant temperature sterilizing after 20 minutes in room temperature preservation, for subsequent use.
Particularly, described enrichment culture is dark condition, is 15-25 DEG C in temperature, and rotating speed is under the condition of 150-200rpm, Secondary Culture 2-3 time, and the Secondary Culture time is 20-30 day/time.
Wherein, the culture condition of described isolation and purification culture is under dark condition, and temperature is 35 DEG C, and incubation time is 2-3 day/time.
The present invention provides the application of described Sphingomonas Sphingomonas sp.815Y in degraded bromate on the other hand, comprise: described Sphingomonas Sphingomonas sp.815Y is inoculated in the substratum containing bromate, carries out shaking culture.
Wherein, the described substratum containing bromate is composed as follows: CaCl
25mg, MgCl
25mg, MnCl
20.3mg, FeCl
20.3mg, ZnCl
20.01mg, H
3bO
30.01mg, CoCl
20.005mg, bromate 1mg, sodium acetate 10mg, distilled water 1,000mL, pH 7.0-8.0, substratum constant temperature sterilizing 20 minutes at 121 DEG C.
Wherein, the culture temperature that described concussion is cultivated is 15-35 DEG C, is preferably 25-35 DEG C.
Particularly, described shaking culture is carried out under the following conditions: dark condition, and culture temperature is 25-35 DEG C, and rotating speed is 150-170rpm, and concussion incubation time is 15-25 days.
Particularly, described shaking culture condition is: dark condition, and culture temperature is 25 DEG C, rotating speed 150rpm, and concussion incubation time is 20 days.
Wherein, the inoculum size of described Sphingomonas bacterial strain is 0.2-0.3g wet cell/50ml substratum.
Another aspect of the invention provides described Sphingomonas bacterial strain Sphingomonas sp.815Y processing the application in the water polluted by bromate, comprise: in the water polluted by bromate, add described Sphingomonas, mix, cultivate 15-25 days.
Wherein, be under dark condition in described culturing process, culture temperature is 25-35 DEG C, cultivates 15-25 days.
Particularly, in culturing process, add the additional carbon of sodium acetate as Sphingomonas.
Wherein, the addition of additional carbon is add 10-200mg sodium acetate in every premium on currency.
Particularly, the inoculum size of described Sphingomonas bacterial strain is 0.2-0.3g wet cell/50ml water.
Further aspect of the present invention provides described Sphingomonas strain Sphingomonas sp.815Y in the application of degraded bromate in drinking water, comprise: described Sphingomonas is inoculated in the tap water containing bromate, carry out Sphingol single-cell cultivation, the bromate in degraded tap water.
Particularly, described Sphingomonas is inoculated in water treatment and crosses in the tap water obtained containing using ozone oxidation processing technique step in title, carry out Sphingol single-cell cultivation, the bromate in degraded tap water.
Wherein, described tap water prepares in the process of tap water in waterworks, containing using ozone to carry out oxidation processing technique to water inlet, and obtained tap water.
Wherein, Sphingol single-cell is cultivated under the following conditions: dark condition, and culture temperature is 25-35 DEG C, and incubation time is 15-25 days.
Particularly, add the additional carbon of sodium acetate as Sphingol single-cell in culturing process, wherein the addition of additional carbon adds 10-200mg in often liter of tap water.
Particularly, the inoculum size of described Sphingomonas bacterial strain is 0.2-0.3g wet cell/50ml tap water.
Sphingomonas bacterial strain of the present invention (Sphingomonas sp.815Y) tool has the following advantages:
1, Sphingomonas strain of the present invention is strong to the degradation capability of bromate, and degradation speed is fast, is that 1000 μ g/L bromates degraded clearance at short notice reaches more than 50% for concentration;
2, Sphingomonas strain of the present invention is for the treatment of tap water in the water of ozonization process, and the by product bromate that ozone oxidation produces is degraded to bromide anion, improves the security of tap water;
3, Sphingomonas strain of the present invention can add in the water polluted by bromate, and bromate is removed in degraded, carries out biological restoration, reduce its carcinogenic toxicity to the water that bromate pollutes.
4, Sphingomonas strain of the present invention can add in ozone-BAC process unit; as the deeply process means of bromate being carried out to biological restoration; method is simple; run and maintenance cost lower, have a good application prospect in the process of mass-producing process ozone disinfection by-product bromate.
Accompanying drawing explanation
Fig. 1 is the bacterium colony figure of Sphingomonas Sphingomonas sp.815Y on substratum;
Fig. 2 is the cellular form figure of Sphingomonas Sphingomonas sp.815Y;
Fig. 3 is bromate and bromide ion concentration variation diagram in substratum in Sphingomonas Sphingomonas sp.815Y culturing process in embodiment 3;
Fig. 4 is bromate and bromide ion concentration variation diagram in substratum in Sphingomonas Sphingomonas sp.815Y culturing process in embodiment 4;
Fig. 5 processes in test example 1 to be tested by the bromate in bromate polluted water;
Fig. 6 processes in test example 2 to be tested by the bromate in bromate polluted water.
Embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage and disadvantage of the present invention will be more clear along with description.But these embodiments are only exemplary, do not form any restriction to scope of the present invention.It will be understood by those skilled in the art that and can modify to the details of technical solution of the present invention and form or replace down without departing from the spirit and scope of the present invention, but these amendments and replacement all fall within the scope of protection of the present invention.
In embodiment, method therefor is ordinary method if no special instructions, and all percentage concentrations are mass percent concentration, and the solvent in all substratum is distilled water.
The separation of embodiment 1 bacterial strain and purifying
1, medium preparing
(1) enrichment medium:
CaCl
25mg, MgCl
25mg, MnCl
20.3mg, FeCl
20.3mg, ZnCl
20.01mg, H
3bO
30.01mg, CoCl
20.005mg, bromate 1mg, sodium acetate 10mg, distilled water 1,000mL, pH 7.0-8.0, substratum constant temperature sterilizing 20 minutes at 121 DEG C.
(2) separation, pure medium:
Yeast extract 0.5g, Tryptones 0.5g, acid hydrolysis casein 0.5g, glucose 0.5g, Zulkovsky starch 0.5g, K
2hPO
40.3g, MgSO
47H
2o 0.05g, Sodium.alpha.-ketopropionate 0.3g, agar 15g, distilled water 1,000mL, pH 7.0-8.0, substratum constant temperature sterilizing 20 minutes at 121 DEG C.
(3) LB solid medium
Tryptones 10g, yeast extract 5g, NaCl 10g, agar 15g, distilled water 1,000mL, pH7.2.Before substratum uses under 15psi high pressure steam sterilizing 20 minutes.
2, phosphoric acid buffer
Phosphate buffered saline buffer is the mixing solutions of Sodium phosphate dibasic and SODIUM PHOSPHATE, MONOBASIC, and pH is 7.2.
In 800ml distilled water, add 8gNaCl, 0.2KCl, 1.44g Sodium phosphate dibasic and 0.24g potassium primary phosphate, regulate PH to 7.2 with hydrochloric acid, add water and be settled to 1L.Under 15psi high pressure, steam sterilizing was stored in room temperature after 20 minutes, for subsequent use.
3, enrichment culture
Ozone-bioactive carbon the water treatment device of the ambient water quality National Key Laboratory environmental microorganism technical study group of Ecological Environment Research Center, Chinese Academy of Sciences, first this treatment unit carries out ozone Oxidation Treatment to water inlet, the bromate of higher concentration is contained through the water of ozone Oxidation Treatment, current are through gac, carry out absorption and biological degradation process, wherein, treatment unit runs 6 months continuously under normal temperature (25 DEG C) condition; Ozone dosage is 2.5-3.0mg/L; The flow velocity of biological activated carbon adsorption column is 7ml/min, and bioactive carbon post cubic capacity is 450ml, and the bromate concentration of water inlet is 10-20 μ g/L, and organic content is 2.5-3.0mg/L, and the bromate of water outlet is 0-5 μ g/L.
Take the biological activated carbon 10g be collected in ozone-bioactive carbon water treatment device, add phosphate buffered saline buffer and little granulated glass sphere, oscillation treatment 4h under 150rpm, obtained bacteria suspension, then carries out centrifugal concentrating process to bacteria suspension, then get 10mL gained and concentrate bacterium liquid, be inoculated in the serum bottle containing 300mL enrichment medium, under dark condition, in 25 DEG C, 150rpm, cultivate 20 days, obtain first-generation nutrient solution; Then getting first-generation nutrient solution 10ml continues to be inoculated in the serum bottle that 300mL enrichment medium is housed, and under dark condition, in 25 DEG C, 150rpm, cultivates 20 days, obtains s-generation nutrient solution; Then getting s-generation nutrient solution 10ml continues to be inoculated in the serum bottle that 300mL enrichment medium is housed, under dark condition, in 25 DEG C, 150rpm, cultivate 20 days, obtain third generation enrichment culture liquid, be about to the concentrated bacterium liquid Secondary Culture 2 times obtained through centrifugal concentrating process, obtain enrichment culture liquid.
3, separation, purifying are cultivated
By separation and Culture of ruling on plate isolation, pure medium after third generation nutrient solution (i.e. enrichment culture liquid) dilution, put into constant incubator and cultivate 5 days at 35 DEG C.The single bacterium colony that flat board under picking appropriate dilution is formed, streak inoculation is in plate isolation, pure medium again, carry out separation and Culture, cultivate at 35 DEG C after 5 days and microscopy is carried out to plate culture, if the culture bacterial classification obtained is impure, then the streak inoculation of picking plate culture continues purifying in plate isolation, pure medium as stated above, until microscopy result is indicated as pure bacterium; Purifiedly obtain pure Sphingomonas.
The Microbiological Characteristics research of embodiment 2 Sphingomonas
1, colony morphological observation
By being separated, purifying cultivates the unit cell bacterial strain streak inoculation that obtains in LB solid medium, and be cultured at 35 DEG C and grow bacterium colony, observation colonial morphology, result is as follows:
The colony characteristics of bacterial strain is: the colony diameter size of cultivating 2 days on LB flat board is about 1-2mm, and bacterium colony is in yellow, moistening, droplet-like, circular, protruding, opaque, neat in edge.As shown in Figure 1.
2, cellular form is observed
Picking cultivates the bacterial strain bacterium colony of 2 days on LB flat board, successively with glutaraldehyde fix, phosphate buffered saline buffer (pH7.2) rinsing, graded ethanol dehydration, isoamyl acetate process and the drying of carbon dioxide critical point, metal spraying, sample is put into observation ward after carry out electron-microscope scanning, electron-microscope scanning result is as Fig. 2, show that Sphingol single-cell is shaft-like, not sprout spore, atrichia, size is 0.53 ~ 0.70 μm × 1.06 ~ 1.67 μm.
3,16S rRNA gene sequencing
By the Zymomonas mobilis that is inoculated in LB substratum in 25 DEG C, Pseudomonad cell after 150rpm cultivates 20 days adopts DNA extraction kit (FastDNA SPIN kit for soil after collecting, MP Biomedicals, France), extract the DNA of Zymomonas mobilis; Adopt forward primer (27f primer, 5 '-CAGGAAACAGCTATGAC-3 ') and reverse primer (1492r primer, 5 '-TACGGYTACC TTGTTACGAC TT-3 ') to the 16SrRNA of bacterial strain in the amplification of PCR electrophoresis apparatus enterprising performing PCR; Amplified production is adopted purification kit (Fermantas, Gene JET Extraction Kit, EU) purifying is carried out, then adopt TA-cloning process (molecular cloning handbook) to be checked order by the intestinal bacteria (TOP10 Competent cell) after successful conversion the PCR primer after purifying, sequencing result is compared in NCBI gene library (http://blast.ncbi.nlm.nih.gov/).
Sequencing result is shown in sequence table, and wherein, 16S rRNA (the SEQ ID NO.1) sequence length of bacterial strain is 1466bp.
According to the colony morphology characteristic of bacterial strain, the comparison result of cell morphological characteristic and its 16S rRNA gene order, qualification bacterial strain belongs to Sphingomonas sp..
Sphingol single-cell mentioned in the present invention is through qualification, find that its strain morphology and physiological and biochemical test result and Sphingomonas koreensis feature are the most close, Sphingomonas koreensis is Gram-negative bacteria, has one pole flagellum, mobility bacillus.Colonial morphology is rounded, dimpling face, smooth surface, opaque in yellow.Find through 16S rRNA sequence alignment, both similaritys are 98%, and therefore Sphingomonas of the present invention is different from existing Sphingol single-cell, up to now without report.
Sphingomonas of the present invention has been preserved in that " China Committee for Culture Collection of Microorganisms's common micro-organisms " center "; its strain name is Sphingol single-cell Sphingomonas sp.815Y, " deposit number of China Committee for Culture Collection of Microorganisms's common micro-organisms " center " is CGMCC No.4721 on March 28th, 2011.
Embodiment 3
1) wet cell of unit cell bacterial strain Sphingomonas sp.815Y is prepared
The fresh slant strains of Sphingomonas Sphingomonas sp.815Y is got with transfering loop, streak inoculation is on LB solid medium, at 35 DEG C, cultivate after 2 days and can obtain the long culture medium flat plate having obvious bacterium colony, obtain fresh Sphingol single-cell wet cell.
2) bromate test is removed
The wet cell of picking bacterial classification Sphingomonas sp.815Y is inoculated in enrichment medium, under dark condition, cultivates in 25 DEG C, wherein, inoculates 0.2g wet cell in every 50mL enrichment medium; Bromate in nutrient solution and ionic bromide content adopt ion-chromatographic determination according to the method for standard GB/T/T 5750.10-2006 (drinking water standard method of inspection disinfection byproduct (DBP) index), measure the concentration value of bromate and bromide anion in the nutrient solution of after inoculating front substratum and inoculating the 9th day, 15 days, 20 days, measurement result as shown in Figure 3.
Measurement result shows: Sphingol single-cell Sphingomonas sp.815Y of the present invention has good removal ability to bromate, high to the removal efficiency of bromate, when inoculum size is 0.2g wet cell/50mL nutrient solution, more than 40% is reached to the clearance of bromate in cultivation within 15 days.
Embodiment 4
Except step 2) in the inoculum size of bacterial classification Sphingomonas sp.815Y be inoculate 0.3g wet cell in every 50mL enrichment medium, measure the nutrient solution before inoculation, inoculate outside the concentration of bromate and bromide anion in the nutrient solution of latter 7th day, 15 days, 25 days, all the other are identical with embodiment 3, and measurement result as shown in Figure 4.
Measurement result shows: Sphingol single-cell Sphingomonas sp.815Y of the present invention has good removal ability to bromate, high to the removal efficiency of bromate, when inoculum size is 0.3g wet cell/50mL nutrient solution, after cultivation 15 days, more than 60% is reached to the clearance of bromate.
Reference examples 1
Except carrying out high pressure steam sterilization immediately by after the wet cell of inoculation Sphingomonas sp.815Y, i.e. steam sterilizing 20 minutes under 15psi high pressure, outside the Sphingomonas sp.815Y deactivation immediately making inoculation, obtain the control group of inactive, all the other are identical with embodiment 3, measure bromate and bromide ion concentration in the corresponding nutrient solution of after inoculating front substratum and inoculating the 9th day, 15 days, 20 days, measurement result as shown in Figure 3.
Measurement result is that bromate in the nutrient solution of reference examples 1 and bromide ion concentration remain unchanged, blank determination result shows: in nutrient solution, the degraded of bromate is caused by Sphingol single-cell, and the biomass after sterilizing to bromate without adsorptive power, therefore, Sphingol single-cell Sphingomonas sp.815Y can remove bromate effectively, can be used for the degraded of bromate.
Reference examples 2
Except carrying out high pressure steam sterilization immediately by after the wet cell of inoculation Sphingomonas sp.815Y, i.e. steam sterilizing 20 minutes under 15psi high pressure, outside the Sphingomonas sp.815Y deactivation immediately making inoculation, obtain the control group of inactive, all the other are identical with embodiment 4, measure the concentration of bromate and bromide anion in the nutrient solution before inoculation, the inoculation nutrient solution of latter 7th day, 15 days, 25 days, measurement result as shown in Figure 4.
Measurement result is that bromate in the nutrient solution of reference examples 2 and bromide ion concentration remain unchanged, blank determination result shows: in nutrient solution, the degraded of bromate is caused by Sphingol single-cell, and the biomass after sterilizing to bromate without adsorptive power, therefore, Sphingol single-cell Sphingomonas sp.815Y can remove bromate effectively, can be used for the degraded of bromate.
Test example 1 removes the bromate test in the tap water of bromate pollution
1) the fresh slant strains of Sphingomonas Sphingomonas sp.815Y is got with transfering loop, streak inoculation, on LB solid medium, at 35 DEG C, is cultivated after 2 days and can be obtained the long culture medium flat plate having obvious bacterium colony, obtain fresh Sphingol single-cell wet cell, for subsequent use.
2) in tap water, add sodium bromate, the bromate concentration of tap water is promoted to about 1100 μ g/L, the pH of tap water is 7.5;
3) add sodium acetate wherein by containing after the tap water autoclaving process of bromate, the addition of sodium acetate is 10mg/L, namely adds sodium acetate 10mg in every 1L sterile tap water;
4) get the tap water 300ml after sterilizing and be placed in 500ml serum bottle, inoculation Sphingol single-cell Sphingomonas sp.815Y, after sealing under dark condition, quiescent culture at temperature 25 DEG C, bromate in degraded tap water, inoculum size is 0.3g wet cell/100ml;
5) adopt latter 0th, 1 day, 4 days of ion-chromatographic determination inoculation, the bromate in drinking water of 6 days and the concentration of bromide anion according to the method for standard GB/T/T 5750.10-2006 (drinking water standard method of inspection disinfection byproduct (DBP) index), measurement result as shown in Figure 5.
Measurement result shows: after inoculating strain Sphingol single-cell Sphingomonas sp.815Y, Sphingol single-cell was degraded bromate from second day, bromate concentration in tap water declines rapidly, degraded product bromide ion concentration raises gradually, the bromate concentration concentration inoculated after 6 days in tap water is reduced to less than 50% of initial concentration, illustrate that the clearance of Sphingol single-cell Sphingomonas sp.815Y to bromate reaches more than 50%, show that Sphingol single-cell Sphingomonas sp.815Y degraded bromate efficiency is high.Test example 2 removes the bromate test in the tap water of bromate pollution
1) the fresh slant strains of Sphingomonas Sphingomonas sp.815Y is got with transfering loop, streak inoculation, on LB solid medium, at 35 DEG C, is cultivated after 2 days and can be obtained the long culture medium flat plate having obvious bacterium colony, obtain fresh Sphingol single-cell wet cell, for subsequent use.
2) in tap water, add sodium bromate, the bromate concentration of tap water is promoted to about 1000 μ g/L, the pH of tap water is 7.5;
3) get the tap water 300ml after sterilizing and be placed in 500ml serum bottle, inoculation Sphingol single-cell Sphingomonas sp.815Y, after sealing under dark condition, quiescent culture at temperature 25 DEG C, bromate in degraded tap water, inoculum size is 0.3g wet cell/100ml;
4 adopt the rear bromate in drinking water of the 0th, 1,2,3,5,6 day of ion-chromatographic determination inoculation and the concentration of bromide anion according to the method for standard GB/T/T 5750.10-2006 (drinking water standard method of inspection disinfection byproduct (DBP) index), and measurement result as shown in Figure 6.
Measurement result shows: after inoculating strain Sphingol single-cell Sphingomonas sp.815Y, Sphingol single-cell from inoculation first day namely to degrade bromate, bromate concentration in tap water declines rapidly, degraded product bromide ion concentration raises gradually, the bromate concentration concentration inoculated after 6 days in tap water is reduced to less than 70% of initial concentration, illustrate that the clearance of Sphingol single-cell Sphingomonas sp.815Y to bromate reaches more than 30%, show that Sphingol single-cell Sphingomonas sp.815Y degraded bromate efficiency is high; But due to not for Zymomonas mobilis provides additional carbon (sodium acetate), the degradation efficiency of bacterial strain to bromate is slightly poorer than the degradation effect that with the addition of Additional carbon sources.
Claims (11)
1. a strain Sphingol single-cell (Sphingomonas sp.) 815Y bacterial strain, it is characterized in that, its microbial preservation is numbered CGMCC No.4721.
2. the application of Sphingomonas strain as claimed in claim 1 in degraded bromate.
3. apply as claimed in claim 2, it is characterized in that, described Sphingomonas strain is inoculated in the substratum containing bromate, carries out Sphingol single-cell cultivation.
4. apply as claimed in claim 3, it is characterized in that, described culture temperature is 15-35 DEG C.
5. Sphingomonas strain as claimed in claim 1 is processing the application in the water polluted by bromate.
6. apply as claimed in claim 5, it is characterized in that, in the water polluted by bromate, add described Sphingomonas, mix, cultivate 15-25 days.
7. Sphingomonas strain as claimed in claim 1 is in the application of degraded bromate in drinking water.
8. apply as claimed in claim 7, it is characterized in that, described Sphingomonas is inoculated in the tap water containing bromate, carries out Sphingol single-cell cultivation, the bromate in degradation water.
9. apply as claimed in claim 8, it is characterized in that described culture temperature is 15-35 DEG C.
10. the application as described in claim 6 or 8, is characterized in that, adds the additional carbon of sodium acetate as Sphingomonas in culturing process.
11. apply as claimed in claim 10, it is characterized in that, the addition of described sodium acetate is add 10-200mg sodium acetate in every premium on currency.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1796314A (en) * | 2004-12-24 | 2006-07-05 | 中国科学院生态环境研究中心 | Method for processing urban sewage deeply |
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| CN1796314A (en) * | 2004-12-24 | 2006-07-05 | 中国科学院生态环境研究中心 | Method for processing urban sewage deeply |
Non-Patent Citations (3)
| Title |
|---|
| Bromate Reduction by Denitrifying Bacteria;W. A. M. HIJNEN et al.;《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》;19950131;第61卷(第1期);239-244 * |
| O3 —BAC工艺的微生物群落结构解析;王振宇等;《中国给水排水》;20060131;第22卷(第1期);1-4 * |
| 含溴水源水臭氧处理时溴酸盐的产生与控制;裴义山等;《环境科学学报》;20071130;第27卷(第11期);1767-1770 * |
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