CN106365310B - A kind of method that clay mineral cooperates with promotion biological reducing nitrobenzene with humic acid - Google Patents
A kind of method that clay mineral cooperates with promotion biological reducing nitrobenzene with humic acid Download PDFInfo
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- CN106365310B CN106365310B CN201610921607.2A CN201610921607A CN106365310B CN 106365310 B CN106365310 B CN 106365310B CN 201610921607 A CN201610921607 A CN 201610921607A CN 106365310 B CN106365310 B CN 106365310B
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- humic acid
- clay mineral
- nitrobenzene
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/28—Anaerobic digestion processes
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/38—Organic compounds containing nitrogen
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- Biodiversity & Conservation Biology (AREA)
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- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
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Abstract
The invention discloses a kind of clay minerals to cooperate with the method for promoting biological reducing nitrobenzene with humic acid, belongs to the biologic treating technique field of pollutant.Clay mineral and humic acid are added into nitrobenzene-containing solution, is transferred in serum bottle, and alienation metal reducing miroorganisms cell is added, is subsequently placed in 150rpm, is protected from light 60h in 30 DEG C of constant-temperature tables.Clay mineral used in the present invention and humic acid are NAu-2 and ESHA respectively, and using it is preceding serum bottle concussion both is made to be uniformly mixed for 24 hours, adsorption equilibrium.This method collaboration uses clay mineral and humic acid, can effectively facilitate nitrobenzene biological reducing, and may be implemented to be used repeatedly, and facilitation effect is stablized, and shows better than the facilitation effect being added separately under the conditions of clay mineral and humic acid.Clay mineral and humic acid are all natural soil component, widely distributed, cheap and easy to get, convenient for promoting and applying in practice.
Description
Technical field
The invention belongs to pollutant biologic treating technique field, provides a kind of application clay pit and humic acid collaboration promotes
The method of micro-reduction nitrobenzene.
Background technique
Nitrobenzene be a hydrogen atom on phenyl ring replaced by nitro and the compound that generates, molecular formula C6H5NO2, again
Name mirbane oil, almond oil.Usually as the original of the organic synthesis such as dyestuff, explosive, rubber, pesticide intermediate or production aniline
Expect and is widely present in the waste water of the industries such as dyestuff, pesticide, medicine, petrochemical industry.Nitrobenzene belongs to organic compound difficult to degrade
Object, chemical property are stablized, and have carcinogenicity and mutagenicity to biology, cause after entering environment to water body and soil persistance
Pollution.Therefore, the pollution control of nitrobenzene waste water is constantly subjected to the concern of people.
Currently, the main path for the treatment of of Nitrobenzene waste water includes bioanalysis, chemical method, physical method and combinations thereof technique.It is micro-
Biology can restore conversion nitrobenzene under anaerobic and generate aniline, and complete mine is then further realized under aerobic condition
Change.
Humic acid as a kind of natural organic matter universally present in natural environment, and be easy to from water body, soil with
And it is extracted in bed mud.Studies have shown that humic acid can promote the extracellular electron transmission of bacterium, micro-reduction nitre is significantly improved
The rate of the pollutants such as base benzene.Liu G etc. was in World Journal of Microbiology and in 2013
The 1723-1730 pages of volume 29 of Biotechnology, Cervantes FJ etc. were in Applied Microbiology& in 2012
The 2671-2679 pages of volume 97 of Biotechnology, Liu G etc. were in Applied Microbiology& in 2011
The 417-424 pages and Luan F etc. of volume 91 of Biotechnology 2010 in Environmental Science&
Technology the 184-190 pages paper delivered of volume 44 all reports under humic acid existence condition, azo dyes, nitro
The biological reducing of the pollutants such as benzene is promoted.
Clay mineral is widely present in soils and sediments, is a kind of important iron-bearing mineral.Different from iron (hydrogen) oxygen
Compound mineral, the ferric iron in the clay minerals such as montmorillonite are present in the interlayer structure of mineral, and can be by alienation metal
The micro-reductions such as reducing bacteria are ferrous iron.Luan F etc. 2015 in Environmental Science&Technology
The 1418-1426 pages of volume 49 addition for reporting ferruginous clay mineral can promote the biological reducing of nitrobenzene.
To sum up, humic acid and ferruginous clay mineral, which each can be used as, strengthens the biological reducing that mediator promotes nitrobenzene.
Luan F etc. 2010 in Environmental Science&Technology the 184-190 pages paper report delivered of volume 44
Road humic acid and the collaboration of ferriferous oxide bloodstone promote biological reducing nitrobenzene.However, what micro-reduction bloodstone generated
Ferrous iron is with Fe2+Form drifts in solution, and easily consumes and is lost, it is difficult to reuse.And biological reducing clay mineral institute
The ferrous iron of generation is primarily present in the layer structure of clay mineral, will not be discharged easily, is expected to be continuously circulated utilization.Root
According to Wang K etc. 2005 in Journal of Environmental Quality the 3420-349 pages opinion delivered of volume 34
It is reported in text, clay pit can adsorb humic acid.Suction-operated by clay pit to humic acid is expected to slow down humic acid stream
It loses, improves the stability of facilitation effect.Currently, there is not yet promoting micro-reduction nitro in relation to humic acid and clay mineral collaboration
The report of benzene.
It is cooperateed with using clay mineral with humic acid and promotes biological reducing nitrobenzene, further enhanced biological reduction nitrobenzene speed
Rate, effect stability can simultaneously be recycled for multiple times.Clay mineral and humic acid are all natural soil component, widely distributed, inexpensive easy
, convenient for promoting and applying in practice.
Summary of the invention
To solve biological treatment nitrobenzene waste water inefficiency and humic acid and Fe2+It is easy to run off, recycles Deng reinforcing mediator
The problems such as utilization rate is low, the present invention provide a kind of side for promoting biological reducing nitrobenzene using clay mineral and humic acid collaboration
Method.Compared to clay mineral and humic acid respectively independent role the case where, the facilitation effect of this method p-nitrophenyl biological reducing
More preferably, and two kinds of substances have widely distributed, cheap and easy to get, the features such as cyclic utilization rate is high.
Technical solution of the present invention:
A kind of method that clay mineral cooperates with promotion biological reducing nitrobenzene with humic acid, steps are as follows:
(A) culture of microorganism: using alienation metal reducing miroorganisms Shewanella oneidensis MR-1 as reduction
The microorganism of nitrobenzene and ferruginous clay mineral；
(B) 10-20mmol/L is added into 10-20mmol/L piperazine-N, N- bis- (2-ethanesulfonic acids) (PIPES) buffer solution
Sodium lactate, sodium lactate are transferred in glass serum vials as electron donor, while adjusting pH to 6.8-7.2；Then pass to nitrogen
The oxygen being aerated into serum bottle head space and aqueous solution drains, and sterilizes, cooling, is transferred to anaerobic state；
(C) under anaerobic state, nitrobenzene is added in serum bottle, ultrasound is uniformly dispersed to nitrobenzene, and nitrobenzene exists
Concentration in mixed solution is 30mg/L；
(D) under anaerobic state, the clay mineral of 10g/L is added in serum bottle, the concentration of clay mineral is 0.1g/
L；The clay mineral is nontronite class clay mineral；
(E) pH of humic acid solution is adjusted to 6.8-7.2 with the NaOH of 1mol/L, and passes through 0.45 μm of membrane filtration,
Obtain filtrate；Under anaerobic state, the humic acid solution after deoxygenation is added to serum bottle, the concentration of humic acid is 50mg/L；
Serum bottle is put into 150rpm, is shaken more than for 24 hours in 30 DEG C of constant-temperature tables, is uniformly mixed clay mineral with humic acid, is adsorbed
Balance；
(F) microbial cell of culture to logarithmic phase latter stage is collected, and is transferred in serum bottle, the dense of microbial cell is made
Degree is 0.25g/L；Then, serum bottle is put into 150rpm, in 30 DEG C of constant-temperature tables, is protected from light 1-3d；
(G) after nitrobenzene reduction is complete, under anaerobic state, adding nitrobenzene to concentration is 30mg/L, probes into clay
Mineral cooperate with the cycle applications effect for promoting biological reducing nitrobenzene with humic acid.
Clay mineral of the present invention is after being ground up, sieved, by mineral grain in 0.01M NaCl solution before using
In be sufficiently stirred, keep its evenly dispersed, while making clay mineral Na+Saturation, improves its absorption property to humic acid.
System, which coexists, the beneficial effects of the invention are as follows clay mineral and humic acid can cooperate with the biology of promotion nitrobenzene also
Original, performance better than clay mineral and humic acid respectively individualism when facilitation effect, and there is good reusing, by force
The biological treatment of p-nitrophenyl is changed.Clay mineral and humic acid used in this method are low in cost, from a wealth of sources, are convenient for
It promotes and applies in practice.
Detailed description of the invention
Fig. 1 be clay mineral and humic acid individually exist and the two concurrent conditions under to the shadow of biological reducing nitrobenzene
It rings.
Fig. 2 is the pseudo-first-order dynamics fitting of biological reducing nitrobenzene in different systems.CtAnd C0: respectively represent t moment and
Nitro phenenyl concentration in zero moment system.
Fig. 3 is experimental group and the initial 12h nitrobenzene reduction rate of each cycle period of control group in reusing experiment
Compare.
Fig. 4 be in reusing experiment experimental group and when each cycle period 60h of control group nitrobenzene reduction degree ratio
Compared with.
Specific embodiment
Below in conjunction with attached drawing and technical solution, a specific embodiment of the invention is further illustrated.
Embodiment 1
Clay mineral NAu-2 is cooperateed with humic acid ESHA promotes micro-reduction nitrobenzene:
(1) culture of microorganism: the bacterium using soya broth culture S.oneidensis MR-1 as reduction nitrobenzene
Kind.High pressure steam sterilization culture medium solution under conditions of 121 DEG C.S.oneidensis MR-1 is connect in aseptic operating platform
It plants to soya broth, inoculative proportion 1:100；Culture medium after inoculation is cultivated for 24 hours in 30 DEG C, 150rpm constant-temperature table,
The S.oneidensis MR-1 bacterium solution for obtaining logarithmic growth phase latter stage is spare.
(2) 10mmol/L PIPES buffer solution is prepared, 10mmol/L sodium lactate is added, and be transferred to 50mL serum bottle
In, while pH is adjusted to 6.8-7.2.Logical nitrogen aeration drains the oxygen in serum bottle head space and aqueous solution, and is put into anaerobism
To ensure anaerobic state in case.
(3) it in anaerobic box, takes 0.03g nitrobenzene to be added in PIPES buffer solution, is settled to 100mL to prepare nitro
Benzene stock solution.By stock solution ultrasonic vibration, nitrobenzene is made to be uniformly dispersed.3mL nitrobenzene stock solution is drawn with liquid-transfering gun, and blood is added
In clear bottle, nitro phenenyl concentration is 30mg/L in reaction system.
(4) it weighs 0.1g clay mineral NAu-2 to be added in 10mL PIPES buffer solution, is sufficiently stirred and clay mineral is made
Stock solution.0.3mL NAu-2 stock solution is shifted in anaerobic box into serum bottle, reaction system CLAY MINERALS AND THEIR SIGNIFICANCE concentration is
0.1g/L。
(5) it weighs 0.03g humic acid ESHA to be dissolved in 60mL water, is sufficiently stirred and humic acid stock solution is made, and adjust
PH to 6.8-7.2.3mL humic acid stock solution is drawn, serum bottle is transferred in anaerobic box, contained humic acid is dense in reaction system
Degree is 50mg/L.Serum bottle is put into 150rpm, is shaken for 24 hours in 30 DEG C of constant-temperature tables, mixes clay mineral with humic acid
It is even, adsorption equilibrium.
(6) the MR-1 cell of culture to logarithmic phase latter stage is collected, and is transferred in serum bottle, bacterial cell concentration is reached
0.25g/L.Then, serum bottle is put into 150rpm, in 30 DEG C of constant-temperature tables, is protected from light 60h.Every 12h sampling is primary, uses
High performance liquid chromatography detects nitro phenenyl concentration variation.All experiments are parallel to be carried out 3 times, while it is thin that only MR-1 is respectively set
Born of the same parents and only addition clay mineral and the biological reducing nitrobenzene control experiment that humic acid is only added.
Fig. 1 is experimental group and control group nitrobenzene reduction result in embodiment 1.The result shows that not adding clay mineral and humic
Acid or the control group that clay mineral is only added, humic acid is only added, nitro phenenyl concentration reduces 7.8,9.8 respectively after reacting 12h
And 13.9mg/L；Nitro phenenyl concentration is basicly stable after reacting 60h, and nitrobenzene removal rate is respectively 66%, 76% and 93%.Clay
The addition of mineral or humic acid can promote the reduction of biological p-nitrophenyl in various degree.And it is coexisted in clay mineral and humic acid
Experimental group, nitro phenenyl concentration reduces 19.2mg/L after reacting 12h, compared to only plus the control group of clay mineral or humic acid
96% and 38% are improved respectively, it is with the obvious advantage.
Fig. 2 is experimental group and the fitting of the pseudo-first-order dynamics of control group biological reducing nitrobenzene in embodiment 1.MR-1 cell
Direct-reduction nitrobenzene and the control group first order reaction rate constants k for being individually added into clay mineral or humic acidobsRespectively 1.8
×10-2h-1(r2=0.972), 4.2 × 10-2h-1(r2=0.983) and 2.6 × 10-2h-1(r2=0.973)；And clay mineral and
The k for the experimental group that humic acid coexistsobsIt is 5.1 × 10-2h-1(r2=0.992), it is respectively 2.8 times, 1.2 times of three control groups
With 2.0 times, reduction reaction rate significantly improves.
The experimental results showed that clay mineral and humic acid coexist system have to S.oneidensis MR-1 reduction nitrobenzene it is bright
Aobvious facilitation, and show and be better than that clay mineral is only added or the biological reducing effect under the conditions of humic acid is only added.
Embodiment 2
The cycle applications for promoting biological reducing nitrobenzene are cooperateed with humic acid using clay mineral.
(1) 10mmol/L PIPES buffer solution is prepared, is transferred in 50mL serum bottle after 10mmol/L sodium lactate is added,
And pH is adjusted to 6.8-7.2.Logical nitrogen aeration drains the oxygen in serum bottle head space and aqueous solution, and be put into anaerobic box with
Ensure anaerobic state.
(2) in anaerobic box, 3mL nitrobenzene stock solution (preparation method is shown in embodiment 1) is drawn with liquid-transfering gun, serum is added
In bottle, nitro phenenyl concentration is 30mg/L in reaction system.
(3) clay mineral stock solution 0.3mL being drawn in anaerobic box and being transferred to serum bottle, reaction system CLAY MINERALS AND THEIR SIGNIFICANCE is dense
Degree is 0.1g/L.
(4) 3mL humic acid stock solution being drawn in anaerobic box and being transferred to serum bottle, humic acid concentration is in reaction system
50mg/L.Serum bottle is then put into 150rpm, is shaken for 24 hours in 30 DEG C of constant-temperature tables, mixes clay mineral with humic acid
It is even, adsorption equilibrium.
(5) microbial cell of culture to logarithmic phase latter stage is collected, and is transferred in serum bottle, bacterial cell concentration is made
0.25g/L。
(6) serum bottle is put into 150rpm, in 30 DEG C of constant-temperature tables, is protected from light 60h.Every 12h sampling is primary, using height
Effect liquid phase chromatogram method detects nitro phenenyl concentration variation.All experiments are parallel to be carried out 3 times, while only MR-1 cell is respectively set
And clay mineral is only added and the biological reducing nitrobenzene control experiment of humic acid is only added.
(7) after experimental group nitrobenzene reduction fully reacting, according to liquid chromatography detect remaining nitro phenenyl concentration,
It is supplemented again in anaerobic box to reaction system and nitrobenzene is added, restore its concentration to 30mg/L.Nitro is measured by sampling every 12h
Benzene concentration.Step (6), (7) are repeated, 3 periods are recycled.
Fig. 3 is the comparison of initial 12h nitrobenzene reduction rate in experimental group and each cycle period of control group in embodiment 2.
The result shows that the initial reduction rate of each cycle biological reduction nitrobenzene of system coexists always than only adding in clay mineral and humic acid
Enter clay mineral and the control group high 64%-94% and 14%-37% of humic acid is only added, repeats facilitation effect and stablize.
Fig. 4 be in embodiment 2 in different cycle period experimental groups and control group when 60h nitrobenzene reduction degree comparison.
The result shows that the reduction rate that system nitrobenzene coexists in each cycle period CLAY MINERALS AND THEIR SIGNIFICANCE and humic acid reaches 80% or more, have
Good reusing.And compare the control group that clay mineral is only added and humic acid is only added, clay mineral and humic acid
It is more significant with the increase of cycle-index that the promotion effect of system p-nitrophenyl reducing degree coexists.
Experiment shows that the system that coexists of clay mineral and humic acid has stable circulation facilitation effect, repeat application
In promotion biological reducing nitrobenzene.
Claims (3)
1. clay mineral cooperates with the method for promoting biological reducing nitrobenzene with humic acid, which is characterized in that steps are as follows:
(A) culture of microorganism: using alienation metal reducing miroorganisms Shewanella oneidensis MR-1 as reduction nitro
The microorganism of benzene and ferruginous clay mineral；
(B) 10-20mmol/L sodium lactate, lactic acid are added into bis- (2-ethanesulfonic acid) buffer solutions of 10-20mmol/L piperazine-N, N-
Sodium is transferred in glass serum vials as electron donor, while adjusting pH to 6.8-7.2；Nitrogen is then passed to be aerated to serum
Oxygen in bottle head space and aqueous solution drains, and sterilizes, cooling, is transferred to anaerobic state；
(C) under anaerobic state, nitrobenzene is added in serum bottle, ultrasound is uniformly dispersed to nitrobenzene, and nitrobenzene is mixing
Concentration in solution is 30mg/L；
(D) under anaerobic state, the ferruginous clay mineral of 10g/L are added in serum bottle, the concentration of ferruginous clay mineral is
0.1g/L；
(E) pH of humic acid solution is adjusted to 6.8-7.2 with the NaOH of 1mol/L, and by 0.45 μm of membrane filtration, obtained
Filtrate；Under anaerobic state, the humic acid solution after deoxygenation is added to serum bottle, the concentration of humic acid is 50mg/L；By blood
Clear bottle is put into 150rpm, shakes more than for 24 hours in 30 DEG C of constant-temperature tables, is uniformly mixed ferruginous clay mineral with humic acid, absorption
Balance；
(F) microbial cell of culture to logarithmic phase latter stage is collected, and is transferred in serum bottle, the concentration of microbial cell is made
0.25g/L；Then, serum bottle is put into 150rpm, in 30 DEG C of constant-temperature tables, is protected from light 1-3d；
(G) after nitrobenzene reduction is complete, under anaerobic state, adding nitrobenzene to concentration is 30mg/L, probes into ferruginous clay
Mineral cooperate with the cycle applications effect for promoting biological reducing nitrobenzene with humic acid.
2. the method according to claim 1, wherein the ferruginous clay mineral are nontronite class ferruginous clay
Mineral NAu-2.
3. method according to claim 1 or 2, which is characterized in that in step (B), leading to nitrogen aeration time is 20min；
Sterilising conditions are 121 DEG C, 20min.
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| CN110655279B (en) * | 2019-10-24 | 2022-03-08 | 江苏方天电力技术有限公司 | Application of humic acid-iron (II) -containing artificial soil compound suspension |
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| JPS6028892A (en) * | 1983-07-28 | 1985-02-14 | Mamoru Uchimizu | Water making process |
| JPS6031892A (en) * | 1983-08-02 | 1985-02-18 | Chiaki Kobayashi | Filter material for purifying sewage |
| CN102233258A (en) * | 2010-04-27 | 2011-11-09 | 安洁士石油技术(上海)有限公司 | Water treatment agent with bentonite being modified by humic acid and preparation method thereof |
| WO2015016238A1 (en) * | 2013-07-31 | 2015-02-05 | 国立大学法人名古屋大学 | Electron transfer system and application for same |
| CN105776579A (en) * | 2016-05-26 | 2016-07-20 | 许婷 | Waste plastic recovery processing sewage treating agent and preparation method thereof |
-
2016
- 2016-10-21 CN CN201610921607.2A patent/CN106365310B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6028892A (en) * | 1983-07-28 | 1985-02-14 | Mamoru Uchimizu | Water making process |
| JPS6031892A (en) * | 1983-08-02 | 1985-02-18 | Chiaki Kobayashi | Filter material for purifying sewage |
| CN102233258A (en) * | 2010-04-27 | 2011-11-09 | 安洁士石油技术(上海)有限公司 | Water treatment agent with bentonite being modified by humic acid and preparation method thereof |
| WO2015016238A1 (en) * | 2013-07-31 | 2015-02-05 | 国立大学法人名古屋大学 | Electron transfer system and application for same |
| CN105776579A (en) * | 2016-05-26 | 2016-07-20 | 许婷 | Waste plastic recovery processing sewage treating agent and preparation method thereof |
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