CN103103147B - 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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CN103103147B
CN103103147B CN201210529569.8A CN201210529569A CN103103147B CN 103103147 B CN103103147 B CN 103103147B CN 201210529569 A CN201210529569 A CN 201210529569A CN 103103147 B CN103103147 B CN 103103147B
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denitrification
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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 M 2012182. 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 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, as one of modal pollutent in water body, not only can bring very big risk to human health, also can cause that body eutrophication destroys the stability of the ecosystem.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.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.Sulphur autotrophic denitrification is the autotrophic denitrification process the most widely of research at present, and it refers to and under anoxic or anaerobic condition, utilizes reduced sulfur (S 2-, S 0, S 2o 3 2-etc.) as electron donor, using nitrate as electron acceptor(EA) simultaneously, be reduced to nitrogen.Visible, sulphur autotrophic denitrification bacterium can be realized when sulphur nitrogen two classes are polluted and being removed, and reaches the object 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 sulphur autotrophic denitrification.But the eighties in 20th century, Robertson and Kuenen isolated first aerobic denitrifying bacteria Paracoccus denitrificans in sulphur removal and the water outlet of denitrification treatment system, had changed people to denitrifying understanding, and provide 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 the NO (main component of nitrogen oxides in effluent, accounts for 95%) that is insoluble in water is transferred to liquid phase from gas phase, thereby not only reduces costs, and can also improve purification efficiency.The Absorption via Chemical Complexation that start to grow up the eighties in 20th century is by liquid absorbent Fe iI(L) and the NO of low solubility there is quick complex reaction, 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 existing in flue gas is easily by Fe iI(L) be oxidized to Fe iII(L), absorption agent was lost efficacy.A kind of both economical effective means is to adopt denitrifying microorganism by Fe iI(L)-NO is reduced to N 2fe regenerates iI(L), adopt simultaneously iron also pathogenic microorganism directly by 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 there is 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 the bacterial strain that heterotrophic denitrification removes nitrate and nitrite in waste water; For the feature of Absorption via Chemical Complexation exhaust gas denitration product, provide a strain under organic carbon source existence condition, can efficiently reduce Fe simultaneously iI(L)-NO and Fe iII(L) bacterial strain, realizes 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, and 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 pH7.2, 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 pH7.2,30 ℃, under the aerobic 150rpm speed conditions of constant temperature, cultivates.
Waste water sulphur autotrophic denitrification synchronized desulfuring denitrogenation nutrient media components: Na 2s 2o 3/ S 0/ Na 2s2～10mM, KNO 3/ KNO 25～15mM, all the other compositions are MgSO 47H 2o5mM, CaCl 25mM, NH 4cl0.28g/L, KH 2pO 40.25g/L, NaHCO 35.4g/L, micro-1mL/L, regulates near pH to 7.2, process sterilising treatment before substratum is used; Waste gas sulphur autotrophic denitrification denitration nutrient media components: Na 2s 2o 3/ S 0/ Na 2s2～10mM, Fe iI(L)-NO2～10mM, all the other compositions are M gsO 47H 2o5mM, CaCl 22H 2o5mM, NH 4cl0.28g/L, KH 2pO 40.25g/L, NaHCO 35.4g/L, micro-1mL/L, regulates near pH to 7.2, process sterilising treatment before substratum is used.
Waste water aerobic denitrification denitrogenation nutrient media components: organic carbon source 3～26g/L, MgSO 47H 2o0.1g/L, Na 2hPO 42H 2o7.9g/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 is used.
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 2o5mM, CaCl 25mM, NH 4cl0.28g/L, KH 2pO 40.25g/L, NaHCO 35.4g/L, micro-1mL/L, regulates near pH to 7.2, process sterilising treatment before substratum is used; 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 2o5mM, CaCl 25mM, NH 4cl0.28g/L, KH 2pO 40.25g/L, NaHCO 35.4g/L, micro-1mL/L, regulates near pH to 7.2, process sterilising treatment before substratum is used.
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 2o5mM, CaCl 25mM, NH 4cl0.28g/L, KH 2pO 40.25g/L, NaHCO 35.4g/L, micro-1mL/L, regulates near pH to 7.2, process sterilising treatment before substratum is used.
Wherein the composition of trace element comprises: EDTA50g/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 for the denitrogenation of waste water sulphur autotrophic denitrification synchronized desulfuring: under anaerobic, using thiosulphate, elemental sulfur or sulfide to reduce nitrate, nitrite as electron donor; This bacterium is used for the denitration of waste gas sulphur autotrophic denitrification: under anaerobic, use Fe iI(L) as complexes absorption, absorb NO, thiosulphate, elemental sulfur or sulfide reduce Fe as electron donor iI(L)-NO.
This bacterium is used for waste water aerobic denitrification denitrogenation: under aerobic condition, the organic carbon source of usining is reduced to nitrogen as electron donor by nitrate or nitrite.
This bacterium is for the denitrogenation of waste water heterotrophic denitrification: under anaerobic, with organic carbon source, as electron donor, reduce nitrate or nitrite; This bacterium is used for the denitration of waste gas heterotrophic denitrification: under anaerobic, use Fe iI(L) as complexes absorption, absorb NO, organic carbon source reduces Fe as electron donor iI(L)-NO.
This bacterium is for waste gas dissimilatory iron reduction regeneration complexes absorption: under anaerobic, the organic carbon source of usining reduces Fe as electron donor iII(L).
The above-mentioned ferrous complexing agent L 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 for 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 alienation reduction complexing iron under the condition that, this bacterial strain can exist at organic carbon source.In denitrogenation of waste water is processed, this bacterium not only can under anaerobic be realized simultaneous desulfurization and denitrification by sulphur autotrophic denitrification, can also by anaerobism heterotrophic denitrification and aerobic denitrification, realize the removal of nitrate or nitrite, and cost is lower, technique is simple, has stronger practical value.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 complexing denitration absorbing liquor in a reaction system iI(L)-NO and Fe iII(L) heterotrophism reduction time, realizes the object that complexing agent regeneration has reached continuous denitration, energy consumption is low, investment and working cost few, non-secondary pollution.
Embodiment
Embodiment 1: bacterial strain of the present invention is for waste water sulphur autotrophic denitrification simultaneous desulfurization and denitrification
Waste water sulphur autotrophic denitrification substratum: MgSO 47H 2o5mM, CaCl 25mM, NH 4cl0.28g/L, KH 2pO 40.25g/L, NaHCO 35.4g/L, KNO 310mM, Na 2s 2o 35H 2o5mM, micro-1mL/L, regulates near pH to 7.2, process sterilising treatment before substratum is used.
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 at 30 ℃ of anaerobic culture boxes, interval certain hour sampling and measuring NO 3 -and S 2o 3 2-the variation of concentration, the results are shown in Table 1.As can be seen from Table 1, the NO that starting point concentration is 10mM 3 -s with 5mM 2o 3 2-, bacterial strain LYM can carry out sulphur autotrophic denitrification process, 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
Embodiment 2: bacterial strain of the present invention is for 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 2o7.9g/L, KH 2pO 41.5g/L, KNO 33g/L, micro-1mL/L, regulates near pH to 7.2, process sterilising treatment before substratum is used.Use aerobic denitrification substratum, under aerobic condition, accessing dry cell weight is the bacterial strain LYM of 0.49 ± 0.01g/L, under 30 ℃ of constant-temperature table 150rpm speed conditions, cultivate, 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 finally to drop to approach zero.
The denitrogenation of table 2 bacterial strain LYM aerobic denitrification
Embodiment 3: bacterial strain of the present invention is for waste gas heterotrophism reduction complexing absorption product Fe iIeDTA-NO
Heterotrophism denitration substratum: MgSO 47H 2o5mM, CaCl 25mM, NH 4cl0.28g/L, KH 2pO 40.25g/L, NaHCO 35.4g/L, glucose 3mM, Fe iIeDTA-NO2.32mM, micro-1mL/L, regulates near pH to 7.2, process sterilising treatment before substratum is used.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 at 30 ℃ of anaerobic culture boxes, interval certain hour sampling and measuring F e iIthe variation of EDTA-NO concentration, the results are shown in Table 3.As can be seen from Table 3, cultivate 6h, F e iIeDTA-NO reduction ratio is 99.6%, F e iIeDTA-NO has obtained good removal.Bacterial strain LYM can utilize the growth of additional organic carbon source under heterotrophism condition, and can reduce F simultaneously e iIeDTA-NO.
Table 3 bacterial strain LYM is to F e iIthe heterotrophism reduction of EDTA-NO
Embodiment 4: bacterial strain of the present invention is for waste gas dissimilatory iron reduction regeneration complexes absorption Fe iIeDTA
Dissimilatory iron reduction substratum: MgSO 47H 2o5mM, CaCl 25mM, NH 4cl0.28g/L, KH 2pO 40.25g/L, NaHCO 35.4g/L, glucose 3mM, Fe iIIeDTA8mM, micro-1mL/L, regulates near pH to 7.2, process sterilising treatment before substratum is used.
Use 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 at 30 ℃ of anaerobic culture boxes, interval certain hour sampling and measuring F e iIthe variation of EDTA concentration, the results are shown in Table 4.As can be seen from Table 4, while there is organic carbon source in substratum, LYM can reduce F e iIIeDTA.At initial F e iIIeDTA concentration is in the dissimilatory iron reduction culture medium culturing base of 8mM, after the cultivation of 10h, and the F in system e iIthe growing amount of EDTA is 6.24mM, F e iIIeDTA reduction ratio reaches 78.0%.
Table 4 bacterial strain LYM is to F e iIIthe alienation reduction of EDTA

Claims (4)

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 secondary coccus LYM apt to change, preserving number: CCTCC M2012182, preservation place: Chinese Typical Representative culture collection center, preservation time: on May 25th, 2012.
2. the cultural method of bacterium described in claim 1, is characterized in that:
(1) under anaerobic, secondary coccus LYM apt to change is inoculated in substratum, near pH7.2,30 ℃, the static cultivation of constant temperature;
Waste water sulphur autotrophic denitrification synchronized desulfuring denitrogenation nutrient media components: Na 2s 2o 3/ S 0/ Na 2s2～10mM, KNO 3/ KNO 25～15mM, all the other compositions are MgSO 47H 2o5mM, CaCl 25mM, NH 4cl0.28g/L, KH 2pO 40.25g/L, NaHCO 35.4g/L, micro-1mL/L, process sterilising treatment before substratum is used; Waste gas sulphur autotrophic denitrification denitration nutrient media components: Na 2s 2o 3/ S 0/ Na 2s2～10mM, Fe iI(L)-NO2～10mM, all the other compositions are MgSO 47H 2o5mM, CaCl 22H 2o5mM, NH 4cl0.28g/L, KH 2pO 40.25g/L, NaHCO 35.4g/L, micro-1mL/L, process sterilising treatment before substratum is used;
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 2o5mM, CaCl 25mM, NH 4cl0.28g/L, KH 2pO 40.25g/L, NaHCO 35.4g/L, micro-1mL/L, process sterilising treatment before substratum is used; 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 2o5mM, CaCl 25mM, NH 4cl0.28g/L, KH 2pO 40.25g/L, NaHCO 35.4g/L, micro-1mL/L, process sterilising treatment before substratum is used;
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 2o5mM, CaCl 25mM, NH 4cl0.28g/L, KH 2pO 40.25g/L, NaHCO 35.4g/L, micro-1mL/L, process sterilising treatment before substratum is used;
(2) under aerobic condition, secondary coccus LYM apt to change is inoculated in substratum, near pH7.2,30 ℃, under constant temperature 150rpm speed conditions, cultivate;
Waste water aerobic denitrification denitrogenation nutrient media components: organic carbon source 3～26g/L, MgSO 47H 2o0.1g/L, Na 2hPO 42H 2o7.9g/L, KH 2pO 41.5g/L, KNO 3/ KNO 21～6g/L, micro-1mL/L, process sterilising treatment before substratum is used.
3. the application of bacterium according to claim 1, is characterized in that:
(1) under anaerobic:
This bacterium is for waste water sulphur autotrophic denitrification nitrogen and desulfurization: usining thiosulphate, elemental sulfur or sulfide reduces nitrate, nitrite as electron donor; This bacterium is used for the denitration of waste gas sulphur autotrophic denitrification: use Fe iI(L) as complexes absorption, absorb NO, thiosulphate, elemental sulfur or sulfide reduce Fe as electron donor iI(L)-NO;
This bacterium is for the denitrogenation of waste water heterotrophic denitrification: with organic carbon source, as electron donor, reduce nitrate or nitrite; This bacterium is used for the denitration of waste gas heterotrophic denitrification: use Fe iI(L) as complexes absorption, absorb NO, organic carbon source reduces Fe as electron donor iI(L)-NO;
This bacterium is for waste gas dissimilatory iron reduction: the organic carbon source of usining reduces Fe as electron donor iII(L) realize complexes absorption regeneration;
(2) under aerobic condition:
This bacterium is used for waste water aerobic denitrification denitrogenation: the organic carbon source of usining is reduced to nitrogen as electron donor by nitrate or nitrite.
4. application according to claim 3, 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).
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