CN103103147A - Mercurial paracoccus strain simultaneously having denitrification and iron reduction functions as well as cultivation method and application of mercurial paracoccus strain - Google Patents

Mercurial paracoccus strain simultaneously having denitrification and iron reduction functions as well as cultivation method and application of mercurial paracoccus strain Download PDF

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CN103103147A
CN103103147A CN2012105295698A CN201210529569A CN103103147A CN 103103147 A CN103103147 A CN 103103147A CN 2012105295698 A CN2012105295698 A CN 2012105295698A CN 201210529569 A CN201210529569 A CN 201210529569A CN 103103147 A CN103103147 A CN 103103147A
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张玉
董西洋
李艳梅
陈明翔
李宁
周集体
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Dalian University of Technology
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Abstract

The invention belongs to the field of environmental biotechnologies, and relates to a mercurial paracoccus strain simultaneously having denitrification and iron reduction functions as well as a cultivation method and an application of the mercurial paracoccus strain. The strain is mercurial paracoccus LYM (Lymphocytotoxicity) and the preservation number of the strain is CCTCC M2012182. The strain can be used for not only realizing synchronous denitrification and desulfuration through sulfur autotrophic denitrification by virtue of wastewater denitrification treatment in an anaerobic condition, but also realizing the removal of nitrate and nitrite through anaerobic heterotrophic denitrification and aerobic denitrification, so that the strain is low in cost and simple in technology and has a strong practical value. By adopting the strain, not only can sulfur autotrophic denitrification denitration be realized in an exhaust gas denitration treatment, but also the synchronous heterotrophic reduction of FeII(L)-NO and FeIII(L) in complexation denitration absorption liquid is realized in a reaction system, so that the regeneration of a complexing agent is realized so as to achieve the purpose of continuous denitration. The cultivation method is low in energy consumption and less in investment and operating cost and has no secondary pollution.

Description

One strain has apt to change secondary coccus bacterial strain, cultural method and the application thereof of denitrification and iron restoring function simultaneously
Technical field
The invention belongs to the Environmental Biotechnology field, be specifically related to apt to change secondary coccus bacterial strain and the purposes in waste water and off gas treatment thereof that a strain has anaerobism sulphur autotrophic denitrification, aerobic denitrification, anaerobism heterotrophic denitrification and dissimilatory iron reduction function.
Background technology
Nitrate not only can bring very big risk to human health as one of modal pollutent in water body, can cause that also body eutrophication destroys the stability of the ecosystem.The denitrification denitrogenation process quilt is thought the most cost-effective nitrate processing mode.Denitrification process is divided into heterotrophic denitrification and autotrophic denitrification according to nutrient type.Most of denitrifying bacterium belongs to heterotroph, need to additionally provide organism as electron donor.But, by oxidation inorganics (H 2, S 2-, S 2O 3 2-, Fe, Fe 2+And NH 4 +Deng) reduce the autotrophic denitrification bacterium of nitrate and be prevalent in equally occurring in nature.The autotrophic denitrification bacterium has unique advantage to low ratio of carbon to ammonium, polluted-water (such as underground water) denitrogenation that nutritive ingredient is barren.The sulphur autotrophic denitrification is the autotrophic denitrification process the most widely of research at present, and it refers to utilize reduced sulfur (S under anoxic or anaerobic condition 2-, S 0, S 2O 3 2-Deng) as electron donor, the while as electron acceptor(EA), is reduced to nitrogen with it with nitrate.As seen, sulphur autotrophic denitrification bacterium can realize removing when sulphur nitrogen two classes are polluted, and reaches the purpose of the treatment of wastes with processes of wastes against one another.
Traditional theory thinks, the denitrification process of bacterium need to just can carry out under an anaerobism or anoxia condition, such as the sulphur autotrophic denitrification.But the eighties in 20th century, Robertson and Kuenen isolated the aerobic denitrifying bacteria Paracoccus denitrificans first in sulphur removal and the water outlet of denitrification treatment system, had changed people to denitrifying understanding, and provided a new thinking for bio-denitrification technology.Generally speaking, the ability that aerobic denitrifying bacteria conforms is strong, growth cycle is shorter, biomass is large and it is lower that dissolved oxygen concentration is required, and denitrification process is thorough and speed is rapid.For this Process of Biological Nitrogen Removal of aerobic denitrification, obtaining a plant height effect aerobic denitrifying bacteria is the most key thing.
" 12 " planning outline is listed oxynitride in the binding indicator system first, and has determined 10% reduction of discharging target, and oxynitride has become the emphasis of China's next stage pollution emission reduction.The difficult point that nitrogen oxides pollution is controlled is how will to be insoluble in the main component of the NO(nitrogen oxides in effluent of water, accounts for 95%) be transferred to liquid phase from gas phase, thus not only reduce costs, can also improve purification efficiency.The Absorption via Chemical Complexation that begin to grow up the eighties in 20th century is by liquid absorbent Fe II(L) and the NO of low solubility quick complex reaction occurs, effectively overcome the mass transfer limit of NO.But the regeneration of complexes absorption directly affects further developing of this technology: 1. Fe II(L) the product Fe after complexed absorption NO II(L)-NO reproduction ratio is more difficult; 2. the oxygen that exists in flue gas is easily with Fe II(L) be oxidized to Fe III(L), make absorption agent lose efficacy.A kind of both economical effective means is to adopt denitrifying microorganism with Fe II(L)-NO is reduced to N 2Fe regenerates II(L), adopt simultaneously iron also pathogenic microorganism directly with Fe III(L) be reduced to Fe II(L).But the Fe with respect to obligate II(L)-NO is pathogenic microorganism and Fe also III(L) go back pathogenic microorganism, search out a kind of Fe that can efficiently reduce simultaneously II(L)-NO and Fe III(L) the more excellent selection of can yet be regarded as of microorganism.
Summary of the invention
The object of the present invention is to provide a strain to have apt to change secondary coccus bacterial strain and the purposes in waste water and off gas treatment thereof of anaerobism sulphur autotrophic denitrification, aerobic denitrification, anaerobism heterotrophic denitrification and dissimilatory iron reduction function.For the character of while sulfur-bearing, nitrogen in inorganic wastewater and waste gas, adopt sulphur autotrophic denitrification bacterium to realize the simultaneous removing of sulphur, nitrogen in waste water and waste gas; Respectively under aerobic and anaerobic conditions, provide a strain can realize that heterotrophic denitrification removes the bacterial strain of nitrate and nitrite in waste water; For the characteristics of Absorption via Chemical Complexation exhaust gas denitration product, provide a strain can efficiently reduce simultaneously Fe under the organic carbon source existence condition II(L)-NO and Fe III(L) bacterial strain is realized absorption agent Fe II(L) regeneration.
The present invention can be achieved in the following manner:
One strain has the apt to change secondary coccus bacterial strain of denitrification and iron restoring function simultaneously, this bacterial strain is named as Paracoccus versutus LYM, on May 25th, 2012 in the center preservation of Chinese Typical Representative culture collection, its preserving number CCTCC NO:M2012182.
The cultural method of this bacterial strain:
Cultural method under secondary coccus anaerobic condition apt to change of the present invention is that secondary coccus LYM apt to change is inoculated in substratum, near pH 7.2, and 30 ℃, the static cultivation of constant temperature anaerobism.
Cultural method under secondary coccus aerobic condition apt to change of the present invention is that secondary coccus LYM apt to change is inoculated in substratum, near pH 7.2,30 ℃, cultivates under the aerobic 150rpm speed conditions of constant temperature.
Waste water sulphur autotrophic denitrification synchronized desulfuring denitrogenation nutrient media components: Na 2S 2O 3/ S 0/ Na 2S 2 ~ 10mM, KNO 3/ KNO 25 ~ 15mM, all the other compositions are MgSO 47H 2O 5mM, CaCl 25mM, NH 4Cl 0.28g/L, KH 2PO 40.25g/L, NaHCO 35.4g/L micro-1mL/L regulates near pH to 7.2, process sterilising treatment before substratum uses; Waste gas sulphur autotrophic denitrification denitration nutrient media components: Na 2S 2O 3/ S 0/ Na 2S 2 ~ 10mM, Fe II(L)-NO 2 ~ 10mM, all the other compositions are MgSO 47H 2O 5mM, CaCl 22H 2O 5mM, NH 4Cl 0.28g/L, KH 2PO 40.25g/L, NaHCO 35.4g/L micro-1mL/L regulates near pH to 7.2, process sterilising treatment before substratum uses.
Waste water aerobic denitrification denitrogenation nutrient media components: organic carbon source 3 ~ 26g/L, MgSO 47H 2O 0.1g/L, Na 2HPO 42H 2O 7.9g/L, KH 2PO 41.5g/L, KNO 3/ KNO 21 ~ 6g/L, micro-1mL/L regulates near pH to 7.2, process sterilising treatment before substratum uses.
Waste water heterotrophic denitrification denitrogenation nutrient media components: organic carbon source 1 ~ 10mM, KNO 3Or KNO 25 ~ 15mM, all the other compositions are MgSO 47H 2O 5mM, CaCl 25mM, NH 4Cl 0.28g/L, KH 2PO 40.25g/L, NaHCO 35.4g/L micro-1mL/L regulates near pH to 7.2, process sterilising treatment before substratum uses; Waste gas heterotrophic denitrification denitration nutrient media components: organic carbon source 1 ~ 10mM, Fe II(L)-NO1 ~ 10mM, all the other compositions are MgSO 47H 2O 5mM, CaCl 25mM, NH 4Cl 0.28g/L, KH 2PO 40.25g/L, NaHCO 35.4g/L micro-1mL/L regulates near pH to 7.2, process sterilising treatment before substratum uses.
Waste gas dissimilatory iron reduction regeneration complexes absorption nutrient media components: organic carbon source 1 ~ 10mM, Fe III(L) 2 ~ 25mM, all the other compositions are MgSO 47H 2O 5mM, CaCl 25mM, NH 4Cl 0.28g/L, KH 2PO 40.25g/L, NaHCO 35.4g/L micro-1mL/L regulates near pH to 7.2, process sterilising treatment before substratum uses.
Wherein the composition of trace element comprises: EDTA 50g/L, ZnSO 422g/L, CaCl 25.54g/L, MnCl 25.06g/L, ammonium molybdate 1.1g/L, CuSO 41.57g/L, CoCl 21.61g/L.
In the present invention, the purposes of secondary coccus LYM apt to change is as follows:
This bacterium is used for the denitrogenation of waste water sulphur autotrophic denitrification synchronized desulfuring: under anaerobic, reduce nitrate, nitrite with thiosulphate, elemental sulfur or sulfide as electron donor; This bacterium is used for the denitration of waste gas sulphur autotrophic denitrification: under anaerobic, use Fe II(L) absorb NO as complexes absorption, thiosulphate, elemental sulfur or sulfide reduce Fe as electron donor II(L)-NO.
This bacterium is used for the waste water aerobic denitrification denitrogenation: under aerobic condition, as electron donor, nitrate or nitrite are reduced to nitrogen with organic carbon source.
This bacterium is used for the denitrogenation of waste water heterotrophic denitrification: under anaerobic, reduce nitrate or nitrite with organic carbon source as electron donor; This bacterium is used for the denitration of waste gas heterotrophic denitrification: under anaerobic, use Fe II(L) absorb NO as complexes absorption, organic carbon source reduces Fe as electron donor II(L)-NO.
This bacterium is used for waste gas dissimilatory iron reduction regeneration complexes absorption: under anaerobic, reduce Fe with organic carbon source as electron donor III(L).
The above-mentioned ferrous complexing agent L that is used for exhaust gas denitration is one or more combinations of ethylenediamine tetraacetic acid (EDTA) (EDTA), nitrilotriacetic acid(NTA) (NTA), hydroxyethylethylene diamine tri-acetic acid (HEDTA), diethylenetriamine pentaacetic acid (DTPA).
The above-mentioned organic carbon source that is used for the heterotrophism process is organic waste or the organic waste water that the organic waste that produces of glucose, sucrose, Sodium.alpha.-hydroxypropionate, methyl alcohol, ethanol, sodium formiate, sodium acetate, Trisodium Citrate, percolate, starch processing or organic waste water, paper industry produce.
The present invention has the following advantages:
Bacterial strain LYM not only can under anaerobic realize heterotrophic denitrification and sulphur autotrophic denitrification, can also under aerobic condition, aerobic denitrification occur.In addition, this bacterial strain can be under the condition that organic carbon source exists alienation reduction complexing iron.In denitrogenation of waste water is processed, this bacterium not only can under anaerobic be realized simultaneous desulfurization and denitrification by the sulphur autotrophic denitrification, can also realize the removal of nitrate or nitrite by anaerobism heterotrophic denitrification and aerobic denitrification, and cost is lower, technique is simple, and stronger practical value is arranged.In exhaust gas denitration is administered, adopt this bacterial strain not only can realize the denitration of sulphur autotrophic denitrification, and Fe in can realizing the complexing denitration absorbing liquor in a reaction system II(L)-NO and Fe III(L) heterotrophism reduction the time realizes that complexing agent regeneration has reached the purpose of continuous denitration, energy consumption is low, investment and working cost few, non-secondary pollution.
Embodiment
Embodiment 1: bacterial strain of the present invention is used for waste water sulphur autotrophic denitrification simultaneous desulfurization and denitrification
Waste water sulphur autotrophic denitrification substratum: MgSO 47H 2O 5mM, CaCl 25mM, NH 4Cl 0.28g/L, KH 2PO 40.25g/L, NaHCO 35.4g/L, KNO 310mM, Na 2S 2O 35H 2O 5mM, micro-1mL/L regulates near pH to 7.2, process sterilising treatment before substratum uses.
Use waste water sulphur autotrophic denitrification substratum, under anaerobic accessing dry cell weight is the bacterial strain LYM of 0.49 ± 0.01g/L, is placed in static cultivation under 30 ℃ of anaerobic culture boxes, interval certain hour sampling and measuring NO 3 -And S 2O 3 2The variation of-concentration the results are shown in Table 1.As can be seen from Table 1, starting point concentration is the NO of 10mM 3 -S with 5mM 2O 3 2-, bacterial strain LYM can carry out sulphur autotrophic denitrification process, through after the cultivation of 72h, and NO 3 -And S 2O 3 2-Clearance be respectively 54.8% and 80.6%.
Table 1 bacterial strain LYM sulphur autotrophic denitrification simultaneous desulfurization and denitrification
Figure BDA00002559237600061
Embodiment 2: bacterial strain of the present invention is used for the waste water aerobic denitrification denitrogenation
Waste water aerobic denitrification substratum: Sodium.alpha.-hydroxypropionate (C 4H 4Na 2O 46H 2O) 13g/L, MgSO 47H 2O0.1g/L, Na 2HPO 42H 2O 7.9g/L, KH 2PO 41.5g/L, KNO 33g/L, micro-1mL/L regulates near pH to 7.2, process sterilising treatment before substratum uses.Use the aerobic denitrification substratum, the access dry cell weight is the bacterial strain LYM of 0.49 ± 0.01g/L under aerobic condition, cultivate under 30 ℃ of constant-temperature table 150rpm speed conditions, the variation of interval certain hour sampling and measuring nitrate nitrogen and nitrite nitrogen concentration the results are shown in Table 2.As can be seen from Table 2, under aerobic condition, through the cultivation of 42h, nearly all nitrate is removed, and nitrite content is first to raise to drop at last to approach zero.
The denitrogenation of table 2 bacterial strain LYM aerobic denitrification
Figure DEST_PATH_GDA00002895924000062
Embodiment 3: bacterial strain of the present invention is used for waste gas heterotrophism reduction complexing absorption product Fe IIEDTA-NO
Heterotrophism denitration substratum: MgSO 47H 2O 5mM, CaCl 25mM, NH 4Cl 0.28g/L, KH 2PO 40.25g/L, NaHCO 35.4g/L, glucose 3mM, Fe IIEDTA-NO 2.32mM, micro-1mL/L regulates near pH to 7.2, process sterilising treatment before substratum uses.Use heterotrophism denitration substratum, under anaerobic accessing dry cell weight is the bacterial strain LYM of 0.49 ± 0.01g/L, is placed in static cultivation under 30 ℃ of anaerobic culture boxes, interval certain hour sampling and measuring Fe IIThe variation of EDTA-NO concentration the results are shown in Table 3.As can be seen from Table 3, cultivate 6h, Fe IIThe EDTA-NO reduction ratio is 99.6%, Fe IIEDTA-NO has obtained good removal.Bacterial strain LYM can utilize under the heterotrophism condition and add organic carbon source growth, and can reduce simultaneously Fe IIEDTA-NO.
Table 3 bacterial strain LYM is to Fe IIThe heterotrophism reduction of EDTA-NO
Figure BDA00002559237600071
Embodiment 4: bacterial strain of the present invention is used for waste gas dissimilatory iron reduction regeneration complexes absorption Fe IIEDTA
Dissimilatory iron reduction substratum: MgSO 47H 2O 5mM, CaCl 25mM, NH 4Cl 0.28g/L, KH 2PO 40.25g/L, NaHCO 35.4g/L, glucose 3mM, Fe IIIEDTA 8mM, micro-1mL/L regulates near pH to 7.2, process sterilising treatment before substratum uses.
Use the dissimilatory iron reduction substratum, under anaerobic accessing dry cell weight is the bacterial strain LYM of 0.49 ± 0.01g/L, is placed in static cultivation under 30 ℃ of anaerobic culture boxes, interval certain hour sampling and measuring Fe IIThe variation of EDTA concentration the results are shown in Table 4.As can be seen from Table 4, when having organic carbon source in substratum, LYM can reduce Fe IIIEDTA.At initial Fe IIIEDTA concentration is in the dissimilatory iron reduction culture medium culturing base of 8mM, through after the cultivation of 10h, and the Fe in system IIThe growing amount of EDTA is 6.24mM, Fe IIIThe EDTA reduction ratio reaches 78.0%.
Table 4 bacterial strain LYM is to Fe IIIThe alienation reduction of EDTA
Figure BDA00002559237600081

Claims (14)

1. a strain has the apt to change secondary coccus bacterial strain of denitrification and iron restoring function simultaneously, it is characterized in that, this bacterium is Paracoccus versutus LYM, preserving number: CCTCC NO:M2012182, preservation place: Chinese Typical Representative culture collection center, preservation time: on May 25th, 2012.
2. the cultural method of the described bacterium of claim 1 is characterized in that: secondary coccus LYM apt to change is inoculated in substratum, and near pH 7.2,30 ℃, the static cultivation of constant temperature anaerobism.
3. the cultural method of the described bacterium of claim 1, is characterized in that: secondary coccus LYM apt to change is inoculated in substratum, near pH 7.2,30 ℃, cultivates under the aerobic 150rpm speed conditions of constant temperature.
4. cultural method according to claim 2 is characterized in that:
Waste water sulphur autotrophic denitrification synchronized desulfuring denitrogenation nutrient media components: Na 2S 2O 3/ S 0/ Na 2S 2 ~ 10mM, KNO 3/ KNO 25 ~ 15mM, all the other compositions are MgSO 47H 2O 5mM, CaCl 25mM, NH 4Cl 0.28g/L, KH 2PO 40.25g/L, NaHCO 35.4g/L, micro-1mL/L, process sterilising treatment before substratum uses;
Waste gas sulphur autotrophic denitrification denitration nutrient media components: Na 2S 2O 3/ S 0/ Na 2S 2 ~ 10mM, Fe II(L)-NO2 ~ 10mM, all the other compositions are MgSO 47H 2O 5mM, CaCl 22H 2O 5mM, NH 4Cl 0.28g/L, KH 2PO 40.25g/L, NaHCO 35.4g/L, micro-1mL/L, process sterilising treatment before substratum uses.
5. cultural method according to claim 3 is characterized in that:
Waste water aerobic denitrification denitrogenation nutrient media components: organic carbon source 3 ~ 26g/L, MgSO 47H 2O 0.1g/L, Na 2HPO 42H 2O 7.9g/L, KH 2PO 41.5g/L, KNO 3/ KNO 21 ~ 6g/L, micro-1mL/L, process sterilising treatment before substratum uses.
6. cultural method according to claim 2 is characterized in that:
Waste water heterotrophic denitrification denitrogenation nutrient media components: organic carbon source 1 ~ 10mM, KNO 3Or KNO 25 ~ 15mM, all the other compositions are MgSO 47H 2O 5mM, CaCl 25mM, NH 4Cl 0.28g/L, KH 2PO 40.25 g/L, NaHCO 35.4g/L, micro-1mL/L, process sterilising treatment before substratum uses;
Waste gas heterotrophic denitrification denitration nutrient media components: organic carbon source 1 ~ 10mM, Fe II(L)-NO 1 ~ 10mM, all the other compositions are MgSO 47H 2O 5mM, CaCl 25mM, NH 4Cl 0.28g/L, KH 2PO 40.25g/L, NaHCO 35.4g/L, micro-1mL/L, process sterilising treatment before substratum uses.
7. cultural method according to claim 2 is characterized in that:
Waste gas dissimilatory iron reduction regeneration complexes absorption nutrient media components: organic carbon source 1 ~ 10mM, Fe III(L) 2 ~ 25mM, all the other compositions are MgSO 47H 2O 5mM, CaCl 25mM, NH 4Cl 0.28g/L, KH 2PO 40.25g/L, NaHCO 35.4g/L, micro-1mL/L, process sterilising treatment before substratum uses.
8. according to claim 4 or 5 or 6 or 7 described cultural methods is characterized in that: the composition of trace element comprises: EDTA 50g/L, ZnSO 422g/L, CaCl 25.54g/L, MnCl 25.06g/L, ammonium molybdate 1.1g/L, CuSO 41.57g/L, CoCl 21.61g/L.
9. the application of bacterium according to claim 1 is characterized in that:
This bacterium is used for waste water sulphur autotrophic denitrification nitrogen and desulfurization: under anaerobic, reduce nitrate, nitrite with thiosulphate, elemental sulfur or sulfide as electron donor; This bacterium is used for the denitration of waste gas sulphur autotrophic denitrification: under anaerobic, use Fe II(L) absorb NO as complexes absorption, thiosulphate, elemental sulfur or sulfide reduce Fe as electron donor II(L)-NO.
10. the application of bacterium according to claim 1 is characterized in that:
This bacterium is used for the waste water aerobic denitrification denitrogenation: under aerobic condition, as electron donor, nitrate or nitrite are reduced to nitrogen with organic carbon source.
11. the application of the described bacterium of claim 1 is characterized in that:
This bacterium is used for the denitrogenation of waste water heterotrophic denitrification: under anaerobic, reduce nitrate or nitrite with organic carbon source as electron donor; This bacterium is used for the denitration of waste gas heterotrophic denitrification: under anaerobic, use Fe II(L) absorb NO as complexes absorption, organic carbon source reduces Fe as electron donor II(L)-NO.
12. the application of the described bacterium of claim 1 is characterized in that:
This bacterium is used for the waste gas dissimilatory iron reduction: under anaerobic, reduce Fe with organic carbon source as electron donor III(L) realize complexes absorption regeneration.
13. according to claim 11 or 12 described application is characterized in that:
Complexing agent L is one or more combinations of ethylenediamine tetraacetic acid (EDTA) (EDTA), nitrilotriacetic acid(NTA) (NTA), hydroxyethylethylene diamine tri-acetic acid (HEDTA), diethylenetriamine pentaacetic acid (DTPA).
14. according to claim 10 or 11 or 12 described application is characterized in that:
The organic carbon source that is used for the heterotrophism process is organic waste or the organic waste water that the organic waste that produces of glucose, sucrose, Sodium.alpha.-hydroxypropionate, methyl alcohol, ethanol, sodium formiate, sodium acetate, Trisodium Citrate, percolate, starch processing or organic waste water, paper industry produce.
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