CN103695349B - A kind of Thermus and application thereof - Google Patents
A kind of Thermus and application thereof Download PDFInfo
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- CN103695349B CN103695349B CN201310697414.XA CN201310697414A CN103695349B CN 103695349 B CN103695349 B CN 103695349B CN 201310697414 A CN201310697414 A CN 201310697414A CN 103695349 B CN103695349 B CN 103695349B
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Abstract
The invention discloses a strain Thermus and purify the application of medium/high sulphur-containing exhaust gas at removing sulphur-containing substance.Water pipe blackening Thermus (Thermus Scotoductus) ZDL1 involved in the present invention, for from Ecological Environment Research Center, Chinese Academy of Sciences's water pollution control laboratory treatment containing the medium/high bioreactor of the sulphur-containing exhaust gas such as sulfur dioxide obtaining separate bacterium, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCC No.7430.This bacterial strain can be prepared as microbial activity filler, for removing the sulfide in medium/high waste gas, purify gas, solve medium/high stench and exhaust pollution problems that the processes such as sewage disposal, sludge anhydration burning, waste incineration and compost produce, the sulphur-containing substance degradation efficiency such as sulfur dioxide, Carbon bisulfide can be reached more than 80%.
Description
Technical field:
The invention belongs to field of environmental improvement, be specifically related to a kind of microorganism removing sulphur-containing substance and application thereof.
Background technology:
During sewage disposal, sludge anhydration burning, compost etc., can produce the waste gas of temperature higher (35～100 DEG C), this medium/high exhaust gas constituents is complicated, and wherein sulfur dioxide is one of main material.Waste gas containing sulfur dioxide spills in air, along with the flowing of air is diffused into everywhere, pollutes surrounding, and health is produced harm.At present, the improvement of waste gas causes people more and more to pay close attention to, and many countries have all formulated relevant discharge standard, the discharge of pollutant in strict control waste gas, and requires enterprise before discharging, takes corresponding control measures.To 33 kinds of atmosphere pollutions in the country " discharge standard of air pollutants " (GB16297-1996) that China came into effect in calendar year 2001, the emission limit of the pollutant such as such as sulfur dioxide is made that strict regulation.
Compared with other physical chemistry waste gas processing method, bioremediation has the advantage such as low cost, easy and simple to handle, technology cleaning, non-secondary pollution because of it, becomes the focus of current research after the nineties.The principle of bioanalysis is that sulphur-containing substance in waste gas is converted into sulfate or the process of sulfur simple substance by the biological oxidation utilizing microorganism.Biological method is suitable to process low concentration, the waste gas of atm number.
But, the most conventional biological treatment technology for waste gas is all to process the material under room temperature, and high-temp waste gas processes the method for reprocessing after generally using spraying cooling water to lower the temperature to waste gas.So, not only increase energy consumption, and need to process newly generated a large amount of waste water, increase processing cost.
Therefore, find the Thermus that can adapt to the existence of medium/high environment, utilize Thermus centering/high-temp waste gas directly to process, it is not necessary to cooling system, thus while reaching exhaust-gas treatment requirement, simplification processes technique, reduces cost.
Summary of the invention:
The problem that the present invention first has to solve is to provide a strain and can directly carry out processing the Thermus removing sulphur-containing substance by centering/high-temp waste gas.
Described Thermus is specially water pipe blackening Thermus (Thermus Scotoductus) ZDL1, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on April 8th, 2013, deposit number is CGMCC No.7430, preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postcode 100101.
This bacterial strain feature is as follows:
This bacterial strain colony colour on LB solid plate is white, opaque, in irregular shape, and the smooth of the edge is moistening.Thalline is elongated rod shape, and size is 0.5 1.5 × 2.5 15.0 μm, atrichia and spore, Gram-negative, catalase and oxidase positive, it is possible to use nitrate, sensitive to penicillin, erythromycin, tetracycline, chloromycetin and streptomycin, but there is anti-polymyxin B and rifampicin characteristic.Optimum growth temperature is 30-100 DEG C.
16SrDNA sequence employing DNAMAN software is carried out similarity merger, it is an operating unit by the similarity sequence merger more than 97%, choosing the representative sequence in each operating unit utilizes the 16SrDNA gene order in BLAST Yu GenBank data base to carry out homology analysis, choose more than 1400bp length to compare (Clustelx1.83), use ortho position to connect (Neighbour Joining) method and carry out Phylogenetic Analysis (MEGA3.1).
Phylogenetic tree is as shown in Figure 1.
To be solved by this invention another problem is that provides water pipe blackening Thermus (Thermus Scotoductus) ZDL1 to purify the application in medium/high sulphur-containing exhaust gas at removing sulphur-containing substance.
Described Thermus ZDL1 is prepared microbial inoculum by the production technology of test tube slant kind-shaking flask kind-seed tank-production tank-product (packaging dosage form is liquid bacterial agent or solid absorption microbial inoculum).
Slant medium consists of: beef powder 3.0-5.0g, Na2S2O3·5H2O5.0g, KH2PO42.0g, KNO32.0g, NH4Cl0.5g, MgCl2·6H2O0.5g, FeSO4·4H2O0.01g, NaHCO31.0g, water 1000ml, pH value 6.0-7.5, agar 15g, 121 DEG C of sterilizing 30min.
Consisting of of liquid fermentation medium: beef powder 3.0-5.0g, Na2S2O3·5H2O5.0g, KH2PO42.0g, KNO32.0g, NH4Cl0.5g, MgCl2·6H2O0.5g, FeSO4·4H2O0.01g, NaHCO31.0g, water 1000ml, pH value 6.0-7.5,121 DEG C of sterilizing 30min.
Described microbial inoculum is 1.0-50m at gas flow3/ h, time of staying 0.5-3min, temperature 50-65 DEG C, sulfur dioxide concentration is 0.1-300ppm, and under the conditions of Carbon bisulfide 0.1-50ppm, the efficiency of the degraded sulphur-containing substance such as sulfur dioxide, Carbon bisulfide can reach more than 80%.Can be used for the sulphur-containing exhaust gas that purification temperature is 50-65 DEG C, solve the medium/high sulphur-containing exhaust gas pollution problem that the processes such as sewage, Treatment of Sludge and compost produce.
Another application of described water pipe blackening Thermus ZDL1 is Thermus ZDL1 appendix to be prepared removing sulphur-containing substance on the porous material purify the microbial activity filler of medium/high sulphur-containing exhaust gas, preferably processes sulphur-containing substance.The filler prepared by this bacterium is 0.001-200m at volume3, the total concentration of sulfide is 30-250ppm, and gas flow rate is 0.05-10000m3/ h, purification temperature is 50-65 DEG C, and under the conditions of time of staying 15s-3min, the degradation efficiency to sulfide reaches more than 85%.
Described porous material is one or more in haydite, granular active carbon, volcanic rock, sponge；
Described porous material density is 0.7-1.0kg/m3, porosity is 50-96%, and particle diameter is 5-50mm；
Described water pipe blackening Thermus ZDL1 is with the form appendix of bacteria suspension on the porous material；
Described water pipe blackening Thermus bacterium ZDL1 suspension is 50-300: 10 with the mass ratio of porous material；
In described Thermus active filler, the cell concentration of water pipe blackening Thermus ZDL1 is more than 106CFU/g( Active filler )。
Described Thermus active filler preparation process is as follows:
(1) slant culture: water pipe blackening Thermus ZDL1 slant strains is prepared in cultivation；
Described water pipe blackening Thermus ZDL1 slant medium consists of: beef powder 3.0-5.0g, Na2S2O3·5H2O5.0g, KH2PO42.0g, KNO32.0g, NH4Cl0.5g, MgCl2·6H2O0.5g, FeSO4·4H2O0.01g, NaHCO31.0g, water 1000ml, agar 15g, pH value 6.0-7.5,121 DEG C of sterilizing 30min.
(2) preparation of bacteria suspension: water pipe blackening Thermus ZDL1 inclined-plane thalline step (1) prepared respectively is by 5-20% inoculum concentration fermentation culture step by step, and separating and obtaining cell concentration is 0.4-0.8g/L water pipe blackening Thermus ZDL1 bacteria suspension；
Water pipe blackening Thermus ZDL1 bacteria suspension preparation method is as follows；
Water pipe blackening Thermus ZDL1 inclined-plane thalline step (1) prepared is by 5-20% inoculum concentration fermentation culture step by step, 60 DEG C of incubation time 48h of cultivation temperature, ventilation 5-200L/min, dissolved oxygen 30%, tank pressure 0.03-0.10Mpa, fermentation liquid is placed in refrigerated centrifuger in 4 DEG C, 2000-8000rpm, centrifugal concentrating 5-20min, abandoning supernatant, precipitation dilutes with sterile nutrient solution respectively, it is thus achieved that cell concentration is 0.4-0.8g/L water pipe blackening Thermus ZDL1 bacteria suspension；
Consisting of of described water pipe blackening Thermus ZDL1 liquid fermentation medium: beef powder 3.0-5.0g, Na2S2O3·5H2O5.0g, KH2PO42.0g, KNO32.0g, NH4Cl0.5g, MgCl2·6H2O0.5g, FeSO4·4H2O0.01g, NaHCO31.0g, water 1000ml, pH value 6.0-7.5,121 DEG C of sterilizing 30min；
Consisting of of described water pipe blackening Thermus ZDL1 sterile nutrient solution: beef powder 3.0-5.0g, KH2PO42.0g, KNO32.0g, NH4Cl0.5g, MgCl2·6H2O0.5g, FeCl2·7H2O0.01g, NaHCO31.0g, water 1000ml, pH value 6.0-7.5,121 DEG C of sterilizing 30min；
(3) preparation of microbial activity filler:
A microbial bacteria suspension prepared by step (2) is sprayed the surface of porous material by ()；B porous material with water pipe blackening Thermus ZDL1 bacterium cell is put into and is rotated mixing in impeller by ()；C () sprays microbial bacteria suspension to porous material surface again, it is thus achieved that microbial activity filler, 4 DEG C save backup；
Preferably repeat step (a)-(c) 2-3 time, make equal appendix in material surface and micropore have bacterium cell, it is thus achieved that microbial activity filler.
Beneficial effect:
1, water pipe blackening Thermus ZDL1 of the present invention, has the efficiency of the higher degraded sulfur-containing foul material such as sulfur dioxide, Carbon bisulfide；Its laundering period is short, can quickly start degradation reaction, applies in the waste gas bioreactor purifying medium/high sulfur-containing foul material, it is possible to shorten the starting period.
2, water pipe blackening Thermus ZDL1 optimum growth temperature of the present invention is 30-100 DEG C, can be used for the sulphur-containing exhaust gas that purification temperature is 30-70 DEG C, the microbial inoculum having this bacterium to prepare can reach more than 80% to the discarded middle sulphur-containing substance degradation efficiency such as sulfur dioxide, Carbon bisulfide, this bacterium the filler prepared is 0.001-200m at volume3, the total concentration of sulfide is 30-250ppm, and gas flow rate is 0.05-10000m3/ h, purification temperature is 50-65 DEG C, and under the conditions of time of staying 15s-3min, the degradation efficiency to sulfide reaches more than 85%.
3, the present invention can be used for the microbial activity filler of preparation removing sulphur-containing substance purification medium/high sulphur-containing exhaust gas, preferably processes sulphur-containing substance.
Accompanying drawing illustrates:
Fig. 1 is water pipe blackening Thermus (Thermus Scotoductus) ZDL1 phylogenetic tree
Detailed description of the invention:
Embodiment 1:
Full-automatic magnetic beads instrument for extracting nucleic acid Smart LabAssist-16 and respective environment sample DNA extraction purification test kit is used respectively purpose bacterial strain DNA to be extracted and purification, selecting bacterial universal primers F16S-27 and R16S-1492 to carry out PCR amplification, primer sequence is respectively as follows:
F16S-27(5`-AGAGTTTGATCCTGGCTCAG-3`)
R16S-1492(5`-CGGTTACCTTGTTACGACTTC-3`)
PCR reaction condition is set as: 94 DEG C of denaturations 5min；94 DEG C of degeneration 30sec subsequently, 56 DEG C of annealing 30sec, 72 DEG C extend 90sec, circulate 25 cycles；Then 72 DEG C extend 10min；Last 4 DEG C keep 10min.Then PCR primer is connected on PMD-19T carrier, is transformed into subsequently in bacillus coli DH 5 alpha competent cell.Competent cell 50 μ l after converting coats in the LB culture medium containing galactoside, isopropyl-β-d-thiogalactoside and ampicillin.After cultivating 12 hours, carry out LB liquid medium cultivation from the single bacterial plaque of each flat board picking 25 white.After cultivating 2 hours, culture fluid is carried out sequencing analysis.
Embodiment 2: use Thermus microbial inoculum to process waste gas
Being centrifuged after water pipe blackening Thermus ZDL1 being fermented and be placed in bioreactor with the microbial inoculum of the resuspended acquisition of nutritional solution, being passed through the gas of sulfur-containing foul material, purifying the sulfur-containing foul material in waste gas by water pipe blackening Thermus ZDL1, Thermus microbial inoculum volume is 1m3, water pipe blackening Thermus ZDL1 content is 1010CFU/ml( Suspension ), the total concentration of sulfide is 150ppm, and gas flow rate is 10m3/ h, purification temperature is 60 DEG C.
Consisting of of described nutritional solution: beef powder 5.0g, KH2PO42.0g, KNO32.0g, NH4Cl0.5g, MgCl2·6H2O0.5g, FeCl2·7H2O0.01g, NaHCO31.0g, water 1000ml, pH value 6.5；The degradation capability of tracing detection sulfur-containing foul material.
The present invention uses gas chromatography tracing detection sulfur-containing compound；Described gas chromatography is: use Agilent GC-6890N chromatographic to measure sulfide concentration, chromatographic column is DB-1701 capillary column (30m × 0.32mm × 0.25 μm), vaporizer, FPD detector, column temperature are respectively set to 100 DEG C, 200 DEG C, 50 DEG C, sample size 100 μ L, N2Flow velocity is 1.7mL/min, and shunt volume is 34.6mL/min.
Result shows that the clearance of sulfide is 82%.
Embodiment 3: a kind of sulphur-containing substance that removes purifies the microbial activity filler of medium/high sulphur-containing exhaust gas
Described porous material appendix has water pipe blackening Thermus ZDL1 bacteria suspension；
Described porous material is granular active carbon；
Described porous material density is 0.8kg/m3, porosity is 86%, and particle diameter is 20mm；
Described water pipe blackening Thermus ZDL1 with the form appendix of bacteria suspension on granular active carbon；
Described water pipe blackening Thermus ZDL1 bacteria suspension is 100: 10 with the mass ratio of granular active carbon；
The cell concentration 5.0 × 10 of water pipe blackening Thermus ZDL1 in described microbial activity filler6CFU/g( Active filler )。
Described microbial activity filler preparation process is as follows:
(1) slant culture: water pipe blackening Thermus ZDL1 slant strains is prepared in cultivation；
Slant medium consists of: beef powder 5.0g, Na2S2O3·5H2O5.0g, KH2PO42.0g, KNO32.0g, NH4Cl0.5g, MgCl2·6H2O0.5g, FeSO4·4H2O0.01g, NaHCO31.0g, water 1000ml, pH value 6.5, agar 15g, 121 DEG C of sterilizing 30min.
(2) preparation of bacteria suspension: water pipe blackening Thermus ZDL1 inclined-plane thalline step (1) prepared respectively is by 10% inoculum concentration fermentation culture step by step, and separating and obtaining cell concentration is 0.6g/L water pipe blackening Thermus ZDL1 bacteria suspension；
Thermus ZDL1 bacteria suspension preparation method is as follows；
Water pipe blackening Thermus ZDL1 inclined-plane thalline step (1) prepared is by 1% inoculum concentration fermentation culture step by step, 60 DEG C of incubation time 48h of cultivation temperature, ventilation 50L/min, dissolved oxygen 30%, tank pressure 0.05Mpa, fermentation liquid is placed in refrigerated centrifuger in 4 DEG C, 6000rpm, centrifugal concentrating 10min, abandoning supernatant, precipitation dilutes with sterile nutrient solution respectively, it is thus achieved that cell concentration is 0.6g/L water pipe blackening Thermus ZDL1 bacteria suspension；
Liquid fermentation medium consist of beef powder 5.0g, Na2S2O3·5H2O5.0g, KH2PO42.0g, KNO32.0g, NH4Cl0.5g, MgCl2·6H2O0.5g, FeSO4·4H2O0.01g, NaHCO31.0g, water 1000ml, pH value 6.0-7.5,121 DEG C of sterilizing 30min；
Consisting of of sterile nutrient solution: beef powder 5.0g, Na2S2O3·5H2O5.0g, KH2PO42.0g, KNO32.0g, NH4Cl0.5g, MgCl2·6H2O0.5g, FeCl2·7H2O0.01g, NaHCO31.0g, water 1000ml, pH value 6.0-7.5,121 DEG C of sterilizing 30min；
(3) preparation of microbial activity filler:
A microbial bacteria suspension prepared by step (2) is sprayed the surface of granular active carbon by ()；B granular active carbon with water pipe blackening Thermus ZDL1 bacterium cell is put into and is rotated mixing in impeller by ()；C () is again to granular active carbon surface spraying microbial bacteria suspension, it is thus achieved that microbial activity filler, 4 DEG C save backup；
Preferably repeat step (a)-(c) 3 times, make equal appendix in material surface and micropore have bacterium cell, it is thus achieved that microbial activity filler.
Claims (3)
1. a Thermus, it is characterised in that: described Thermus be water pipe blackening Thermus (Thermus Scotoductus) ZDL1, deposit number is CGMCC No.7430.
A kind of Thermus the most according to claim 1, it is characterised in that: described water pipe blackening Thermus (Thermus Scotoductus) ZDL1 degraded sulphur-containing substance efficiency reach more than 80%；Described sulphur-containing substance is sulfur dioxide and/or Carbon bisulfide.
A kind of Thermus the most according to claim 1 application in purifying medium/high sulphur-containing exhaust gas, it is characterised in that described sulphur-containing exhaust gas is containing in sulfur dioxide, Carbon bisulfide
The waste gas of at least one；Described medium/high is 50-65 DEG C.
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| CN107937299B (en) * | 2017-11-06 | 2021-02-02 | 福建农林大学 | Thermophilic Pb mineralization bacteria and method for passivating Pb in sludge high-temperature compost by using same |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103084059A (en) * | 2011-11-08 | 2013-05-08 | 财团法人交大思源基金会 | Biological compounds and methods for reducing hydrogen sulfide |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103084059A (en) * | 2011-11-08 | 2013-05-08 | 财团法人交大思源基金会 | Biological compounds and methods for reducing hydrogen sulfide |
Non-Patent Citations (2)
| Title |
|---|
| Identification of iron-reducing Thermus strains as Thermus scotoductus;D.L.Balkwill et al.;《Extremophiles》;20031011;第8卷;第37-44页 * |
| Isolation and characterization of a mixotrophic sulfur-oxidizing Thermus scotoductus;Skirnisdottir S et al.;《Extremophiles》;20010228;第5卷(第1期);第45-51页，尤其是摘要 * |
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