CN105413094B - A kind of charcoal preparation that can mediate pentachlorophenol anaerobic reductive dechlorination - Google Patents
A kind of charcoal preparation that can mediate pentachlorophenol anaerobic reductive dechlorination Download PDFInfo
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- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
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- A62D3/00—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances
- A62D3/02—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances by biological methods, i.e. processes using enzymes or microorganisms
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- A—HUMAN NECESSITIES
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- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
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Abstract
The invention discloses a kind of charcoal preparations that can mediate pentachlorophenol anaerobic reductive dechlorination, are mixed by following components：Minimal medium 1L, 0.5 1mg of anaerobe, 0.6 0.8g of sodium formate, 1 2mL of vitamin solution, 4 6g of Caragana Microphylla biological charcoal.Raw material sources of the present invention are easy to get, and Product environment is friendly, non-toxic, can effectively mediate pentachlorophenol anaerobic reductive dechlorination, the degradation for pentachlorophenol provides new approach.
Description
Technical field
The present invention relates to organic matter biodegradation field, more particularly to a kind of pentachlorophenol anaerobic reductive dechlorination that can mediate
Charcoal preparation.
Background technology
Halogenated aromatic compound and its derivative are that the widely applied chemical industry of the industries such as chemical industry, medicine, process hides, electronics is former
Material, organic synthesis intermediate and organic solvent.Since its pollution is wide, toxicity is big, not degradable, and human body can be generated mostly
Teratogenesis, carcinogenic, mutagenesis " three cause " effect, so as to cause extensive concern.Wherein, pentachlorophenol
（Pentachlorophenol, PCP）It is a kind of typical halogenated aromatic compound, once as insecticide, timber preservative
It is widely used with mould inhibitor etc., so as to which worldwide soil and groundwater be caused to pollute.Since PCP is to human health
（To liver and kidney tool high toxicity and there is initiation cancer equivalent risk）Bring serious threat with ecological environment, thus in
Last century the eighties are prohibited from using.At present, PCP is still identified as persistence organic pollutant, China, the U.S. and the world
Health tissues are classified as priority pollutants.
The structural stability and refractory organics of chloro organic cpd molecule are determined by its chlorination degree, in general
Chlorination degree is higher, and molecular structure is more stable, is also just more difficult to be degraded.Most of chlorinated aromatic organic compounds are come
It says, the distich R-Cl/R-H of standard oxidationreduction potential is between the mV of 250 mV ~ 600.Therefore, these compounds are not
It is easily degraded by Aerobic Process for Treatment, can but carry out reduction conversion under anaerobic.Therefore, microbiological anaerobic reduction dechlorination is
The first step of PCP degradations is also a most important step.
Under the conditions of inventor in microculture system the study found that must have existing for soil or deposit, ability
Carry out the anaerobic reductive dechlorination reaction of pentachlorophenol（Zhang, C.; Suzuki, D.; Li, Z.; Ye, L.;
Katayama, A., Polyphasic characterization of two microbial consortia with
wide dechlorination spectra for chlorophenols. Journal Of Bioscience And Bioengineering 2012, 114, (5), 512-517. Zhang, C.; Katayama, A., Humin as an
Electron Mediator for Microbial Reductive Dehalogenation. Environmental Science & Technology 2012, 46, (12), 6575-6583.）.Pass through consulting literatures, it has been found that this feelings
Condition is not special case.There are many researchs to report that certain dehalogenate floras need to rely on the presence of soil or deposit at present,
To keep their degrading activity.This phenomenon is micro- the halogenated organic pollution such as Polychlorinated biphenyls, dioxin and tetrabromobisphenol A
It is showed in biodegradation particularly typical.Under anaerobic, the reduction dechlorination reaction of most of halogenated organic pollution be all
It has an effect in soil or deposit.For a long time, soil or deposit rise during micro-reduction dehalogenate
Specific effect never has breakthrough.In a recent study, the humin that we are reported in soils and sediments is
Promote micro-reduction dehalogenation（Pentachlorophenol dechlorination and tetrabromobisphenol A debrominate）The necessary factor of generation（Zhang,
C.; Li, Z.; Suzuki, D.; Ye, L.; Yoshida, N.; Katayama, A., A humin-dependentDehalobacter species is involved in reductive debromination of
tetrabromobisphenol A. Chemosphere 2013）.Humin plays electronic media in this reaction process
The effect of body.That is, organic pollution, as why the anaerobic biodegradation activity of PCP can only be in soil or deposit
In reacted, the humin this is because functional microorganism needs to rely in soils and sediments carries out electron transmission,
So as to achieve the effect that dechlorination.
Humin is the solid organic matters for being both also insoluble in alkali in soil or deposit humus insoluble in acid.At present, it learns
It is all water-soluble substance that the electron mediator that persons generally study, which includes humic acid and its pattern compound, in the reality of pollutant
It is easy to be lost in during ground is biological prosthetic.And humin is as solid electronic mediator, it is all insoluble in any pH environment, and its
Chemical constitution is extremely stable, can stay active for long periods.Meanwhile humin or naturally occurring, environmental-friendly, avirulent object
Matter has very strong application advantage in pollution amelioration on the spot.
But the extracting method of humin is troublesome and time-consuming, extraction and purifying every time needs one or two month.
Also, the electronic media effect of humin is often limited by soil or sediment source.
Invention content
The purpose of the present invention is to provide a kind of charcoal preparation that can mediate pentachlorophenol anaerobic reductive dechlorination, raw material comes
Source is easy to get, and Product environment is friendly, non-toxic, can effectively mediate pentachlorophenol anaerobic reductive dechlorination, and the degradation for pentachlorophenol carries
New approach is supplied.
The technical solution adopted by the present invention to solve the technical problems is：
A kind of charcoal preparation that can mediate pentachlorophenol anaerobic reductive dechlorination, is mixed by following components：Inorganic salts
Culture medium 1L, anaerobe 0.5-1mg, sodium formate 0.6-0.8g, vitamin solution 1-2mL, Caragana Microphylla biological charcoal 4-6g.
By studying for a long period of time, inventor is found that a kind of can compare favourably with the effect of the electronic media of humin is imitated in other words
The even better solid electronic mediator-Caragana Microphylla biological charcoal of fruit.The present invention reports Caragana Microphylla biological charcoal as solid electronic matchmaker for the first time
Mediator is in microbial degradation organo-chlorine pollutant（Pentachlorophenol）During play indispensable effect.In view of caragana microphylla is given birth to
Object charcoal is naturally to be made, is environmental-friendly and be widely present in natural environment, the hair of the extracellular electron transport system of this solid
Show, be organic pollution, particularly persistence organic pollutant（Some organic pollutants are easier to be degraded in the environment,
It can be degraded such as under the action of aerobic bacteria or only need the very short time that can be degraded.And as PCP, Polychlorinated biphenyls etc.
Organic matter is difficult to be degraded, and understands and persistently exists in the environment, therefore the microbial degradation utilization of these substances needs specific item
Part need to can just be completed by the Caragana Microphylla biological charcoal in specific carrier such as this patent）The weight of biology in situ recovery technique research
Quantum jump.
The present invention promotes the bio anaerobic reduction dechlorination of pentachlorophenol by the use of Caragana Microphylla biological charcoal as solid electronic mediator.Lemon
Item, big shrub forage plant of falling leaves, root system is extremely flourishing, and main root buries depth, and plant height is 40-70 centimetres, reaches as high as 2 meters of left sides
It is right.Drought-enduring, cold-resistant, high temperature resistant, be aridity grass land, desert steppe area the raw shrubbery of drought.It is NW China, North China, northeast west
One of important species of portion's water and soil conservation and dune-fixating forestation belong to excellent and fix the sand and planting trees on barren hills plant, good forage grass and feed.
Promote the effect of the bio anaerobic reduction dechlorination of pentachlorophenol as solid electronic mediator based on Caragana Microphylla biological charcoal,
Based on Caragana Microphylla biological charcoal, the present invention is specific to have prepared the charcoal preparation that can mediate pentachlorophenol anaerobic reductive dechlorination, packet
Minimal medium, anaerobe, sodium formate and vitamin solution are included, especially can promote the bio anaerobic of pentachlorophenol also
The specific anaerobe selection of former dechlorination.The anaerobic reductive dechlorination of PCP is an electronics consumption process, and microorganism is made by breathing
To organic acid（Such as formic acid）For electron donor, PCP is the process that electron acceptor removes chlorine element from organic pollution, so
Sodium formate in inventive formulation is added in as electron donor.
Preferably, the composition of the minimal medium is：NH4Cl 1.0g/L, CaCl2·2H2O 0.05g/L,
MgCl2·6H2O 0.1g/L, K2HPO4 0.4g/L, SL-10 trace element solution 1mL/L, Se/W solution 1mL/L, pH7.2's
MOPs buffer solution 15mmol/L, water surplus.
Preferably, the composition of every liter of SL-10 trace element solution is：HCl 10.0mL, FeCl2·4H2O 1.5g,
ZnCl270.0mg MnCl2·4H2O 100.0mg, H3BO36.0mg, CoCl2·6H2O 196.0mg, CuCl2·2H2O
2.0mg, NiCl2·6H2O 24.0mg, Na2MoO4·2H2O 36.0mg, water surplus.
Preferably, the composition of every liter of Se/W solution is：NaOH 0.5 g, NaSeO3·5H2O 3 mg, Na2WO4·2H2O
4 mg, water surplus.
Preferably, by mass percentage, the anaerobe is mixed by following strain：Sulphur monad
38-40%, bacteroid 31-32%, Clostridium bifermentans 19-20%, dehalogenate bacillus 4-6%, desulfovibrio 4-6%.The present invention
Specific anaerobe composition can effectively facilitate the bio anaerobic reduction dechlorination of pentachlorophenol.
Preferably, the composition of every liter of vitamin solution is：20 mg of biotin, folic acid 20 mg, VB6100 mg, VB1
50 mg, VB250 mg, VB350 mg, D-VB5 calcium 50 mg, V B12 50 mg, 50 mg of p-aminobenzoic acid, lipoic acid 50
Mg, VB550 mg, 50 mg of heme, 1,2- naphthoquinones, 50 mg, water surplus.
Preferably, the preparation method of the Caragana Microphylla biological charcoal is：Caragana microphylla is cut into the branch of 5-10cm long, in retort
In, under oxygen free condition, 400-500 DEG C of temperature carbonizes 4-6h, takes out, Mechanical Crushing, grinding, you can obtains Caragana Microphylla biological charcoal.
The beneficial effects of the invention are as follows：Raw material sources are easy to get, and Product environment is friendly, non-toxic, can effectively mediate pentachlorobenzene
Phenol anaerobic reductive dechlorination, the degradation for pentachlorophenol provide new approach.
Description of the drawings
The degradation results figure that Fig. 1 is Caragana Microphylla biological charcoal and anaerobe adds in jointly.
The degradation results figure that Fig. 2 is Caragana Microphylla biological charcoal or anaerobe is individually added into.
Specific embodiment
Below by specific embodiment, and with reference to attached drawing, technical scheme of the present invention is described in further detail.
In the present invention, if not refering in particular to, used raw material and equipment etc. are commercially available or commonly used in the art.
Method in following embodiments is the conventional method of this field unless otherwise instructed.
The anaerobe of the present invention is commercially available strain, sulphur monad（Sulfurospirillum halorespiransDSM 13726 ）, bacteroid（Bacteroides graminisolventsDSM 19988）, double enzymes
Clostridium（Clostridium bifermentansATCC 638 ）, dehalogenate bacillus（Dehalobacter restrictusDSM 9455）, desulfovibrio（Desulfovibrio marrakechensisDSM 19337）.Wherein DSM
13726th, DSM 19988, DSM 9455 and DSM19337 are Germany Microbiological Culture Collection Center (DSMZ）Strain number；
ATCC 638 is the number of american strain collection.
The ingredient of minimal medium of the present invention, the ingredient of SL-10 trace element solutions, the ingredient of Se/W solution, dimension life
The ingredient of plain solution is commercial product.
The MOPS buffers of pH7.2 are：41.8 g MOPS are dissolved in 1L distilled water, add the NaOH of 0.5 ~ 1M
Aqueous solution adjusts pH to 7.2, is then settled to 1L with distilled water, in 4 degrees Celsius of preservations.
The preparation method of Caragana Microphylla biological charcoal of the present invention is：Caragana microphylla is cut into the branch of 5-10cm long, in retort, anaerobic
Under the conditions of, 400-500 DEG C of temperature carbonizes 4-6h, takes out, Mechanical Crushing, grinding, you can obtains Caragana Microphylla biological charcoal.
Embodiment 1：
A kind of charcoal preparation that can mediate pentachlorophenol anaerobic reductive dechlorination, is mixed by following components：Inorganic salts
Culture medium 1L, anaerobe 0.5mg, sodium formate 0.6g, vitamin solution 1mL, Caragana Microphylla biological charcoal 4g.
By mass percentage, the anaerobe is mixed by following strain：Sulphur monad 38%, bacteroid
32%, Clostridium bifermentans 20%, dehalogenate bacillus 4%, desulfovibrio 6%.
The composition of minimal medium is：NH4Cl 1.0g/L, CaCl2·2H2O 0.05g/L, MgCl2·6H2O 0.1g/
L, K2HPO4 The MOPs buffer solutions 15mmol/ of 0.4g/L, SL-10 trace element solution 1mL/L, Se/W solution 1mL/L, pH7.2
L, water surplus.
The composition of every liter of SL-10 trace element solution is：HCl （7.7M）10.0mL FeCl2·4H2O 1.5g, ZnCl2
70.0mg MnCl2·4H2O 100.0mg, H3BO36.0mg, CoCl2·6H2O 196.0mg, CuCl2·2H2O 2.0mg,
NiCl2·6H2O 24.0mg, Na2MoO4·2H2O 36.0mg, water surplus.
The composition of every liter of Se/W solution is：NaOH 0.5 g, NaSeO3·5H2O 3 mg, Na2WO4·2H24 mg of O, water
Surplus.
The composition of every liter of vitamin solution is：20 mg of biotin, folic acid 20 mg, VB6100 mg, VB150 mg,
VB250 mg, VB350 mg, D-VB5 calcium 50 mg, V B12 50 mg, 50 mg of p-aminobenzoic acid, 50 mg of lipoic acid,
50 mg of VB5,50 mg of heme, 1,2- naphthoquinones, 50 mg, water surplus.
Embodiment 2：
A kind of charcoal preparation that can mediate pentachlorophenol anaerobic reductive dechlorination, is mixed by following components：Inorganic salts
Culture medium 1L, anaerobe 1mg, sodium formate 0.8g, vitamin solution 2mL, Caragana Microphylla biological charcoal 6g.
By mass percentage, the anaerobe is mixed by following strain：Sulphur monad 40%, bacteroid
31%, Clostridium bifermentans 19%, dehalogenate bacillus 6%, desulfovibrio 4%.
The other the same as in Example 1.
Embodiment 3：
A kind of charcoal preparation that can mediate pentachlorophenol anaerobic reductive dechlorination, is mixed by following components：Inorganic salts
Culture medium 1L, anaerobe 0.8mg, sodium formate 0.69g, vitamin solution 1mL, Caragana Microphylla biological charcoal 5g.
By mass percentage, the anaerobe is mixed by following strain：Sulphur monad 39.2% intends bar
Bacterium 31.7%, Clostridium bifermentans 19.5%, dehalogenate bacillus 4.8%, desulfovibrio 4.8%.
The other the same as in Example 1.
Degrading experiment：
20mL minimal mediums and 0.1g Caragana Microphylla biological charcoals are added in the serum bottle of 60mL（Final concentration is exactly 5g/L）,
Later toward rushing nitrogen in bottle（Purity is 99.99%.The degradation of PCP is completed under anaerobic.It is for handle to rush nitrogen
Oxygen in culture medium is driven away, prepares anaerobic culture medium）, about 30 minutes rubber stoppers for having polytetrafluorethylecoatings coatings in rear plug
（Teflon-coated butyl rubber stoppers, internal diameter 13mm, the production of wheaton companies of the U.S..Specially using having
The rubber stopper of polytetrafluorethylecoatings coatings is because General Purpose Rubber plug can adsorb the organic matter of PCP etc., is coated on rubber stopper poly-
Tetrafluoroethene can prevent the absorption of PCP, and experimental result is made more preferably to explain）, then cover aluminium lid and seal, finally at 121 DEG C
Under the conditions of moist heat sterilization 30 minutes.
Carry out degradation experiment when, toward the bottle of above-mentioned bacterium of having gone out in pass through 0.2 μm of molecular filter（Use purpose：Just in case dimension life
There is microorganism growth in plain solution and reducing agent solution, filter membrane can prevent it from entering in cultivating system）Add in 0.02mL dimension lifes
Plain solution and 2mmol/L Na2S·9H2O（Reducing agent, it is ensured that the anaerobic condition in culture medium.Prepare 200mmol/L's in advance
Na2S·9H2O storing solutions are stored in 4 degree of refrigerators after high pressure sterilization.200 μ l to 20ml is taken to cultivate from storing solution during degradation experiment
In matrix system, so that its final concentration of 2mmol/L）, then add in 20 μM of PCP（PCP is dissolved in the form of PCP-Na salt
In acetone, what is matched is the storing solution of 5mmol/L.It is taken in 80 μ l to 20mL culture medium systems from storing solution when doing degradation experiment,
So that its final concentration of 20 μm of ol/L）With 10mmol/L sodium formates（Sodium formate solution is that water is dissolved in the form of sodium formate
In, what is matched is the storing solution of 1mol/L, prior high pressure sterilization.200 μ l are taken to 20ml culture mediums from storing solution during degradation experiment
In system, so that its final concentration of 10 mmol/L）, finally access 0.02mg anaerobe.
Above-mentioned culture puts the quiescent culture in dark surrounds at 30 DEG C in the incubator.After culture at least 10 days,
1ml samples are taken from cultivating system with disposable syringe, and therefrom extract using acetonitrile and ethyl acetate as extractant PCP and
Its metabolite, then with the QP5050 gas phases for being equipped with DB-5MS capillary columns (J＆W Scientific, Folsom, CA)
Chromatograph-mass spectrometer (Shimadzu, capital of a country, Japan) carries out sample analysis.
Degrading experiment is divided into three groups：
1st group：Add in Caragana Microphylla biological charcoal+anaerobe（Minimal medium+Caragana Microphylla biological charcoal+anaerobe+dimension
Raw element solution+sodium formate+PCP）；2nd group：Caragana Microphylla biological charcoal is added in, is not added with anaerobe（Minimal medium+caragana microphylla life
Object charcoal+vitamin solution+sodium formate+PCP）；3rd group：Caragana Microphylla biological charcoal is not added with, adds anaerobe（Minimal medium+dimension
Raw element solution+sodium formate+PCP）.All experimental groups are all provided with Duplicate Samples, and every group of experiment is at least in triplicate.
Experimental result：
Fig. 1 shows the situations of the 1st group of experiment, and by above-mentioned cultivating system culture 15 days, pentachlorophenol all realized anaerobism
Reduction dechlorination, generation phenol are final metabolite, i.e. phenol content reaches maximum value.In the cultivating system of 20ml at the 15th day
In add in 20 μM of PCP again, after continuing culture 30 days, pentachlorophenol all realizes anaerobic reductive dechlorinations, and content is reduced to 0 μ
M, while final product phenol content reaches maximum value.Continued to add in pentachlorophenol into cultivating system at the 45th day, experimental result
PCP is completely reduced dechlorination again after two weeks for display, shows that the anaerobic reductive dechlorination activity of this microorganism system is very stable.Fig. 2
Represent the 2nd group, the 3rd group of control experiment is right as a result, find in the presence of individually having Caragana Microphylla biological charcoal or anaerobe
Pentachlorophenol does not almost have degradation effect, and after cultivating 15 days, 25 days, 38 days respectively, the content of pentachlorophenol is basically unchanged.This experiment
Illustrate that Caragana Microphylla biological charcoal can mediate the anaerobic reductive dechlorination of pentachlorophenol to act on really, without condition existing for Caragana Microphylla biological charcoal
Under, the dechlorination reaction of pentachlorophenol without.
In addition, the present invention also uses other different charcoals（Rubbish biological charcoal, bamboo charcoal, corncob charcoal etc.）Instead of caragana microphylla
Charcoal carries out degradation experiment, does not almost have degradation effect to pentachlorophenol.Speculate, Caragana Microphylla biological charcoal may contain other biological charcoal
The active group of unexistent tool electron transfer capacity, which can be under the action of microorganism by redox in culture systems
The electronics generated in the process is transferred to pentachlorophenol, so as to mediate the anaerobic organism reduction dechlorination of pentachlorophenol.
Embodiment described above is a kind of preferable scheme of the present invention, and not the present invention is made in any form
Limitation also has other variants and remodeling under the premise of without departing from the technical solution recorded in claim.
Claims (4)
1. a kind of charcoal preparation that can mediate pentachlorophenol anaerobic reductive dechlorination, which is characterized in that mixed and made by following components
Into：Minimal medium 1L, anaerobe 0.5-1mg, sodium formate 0.6-0.8g, vitamin solution 1-2mL, Caragana Microphylla biological charcoal
4-6g；
The composition of the minimal medium is：NH4Cl 1.0g/L, CaCl2·2H2O 0.05g/L, MgCl2·6H2O 0.1g/
L, K2HPO4 The MOPs buffer solutions 15mmol/ of 0.4g/L, SL-10 trace element solution 1mL/L, Se/W solution 1mL/L, pH7.2
L, water surplus；
By mass percentage, the anaerobe is mixed by following strain：Sulphur monad 38-40%, bacteroid
31-32%, Clostridium bifermentans 19-20%, dehalogenate bacillus 4-6%, desulfovibrio 4-6%；
The composition of every liter of vitamin solution is：20 mg of biotin, folic acid 20 mg, VB6100 mg, VB150 mg, VB2 50
Mg, VB350 mg, D-VB5 calcium 50 mg, V B1250 mg, 50 mg of p-aminobenzoic acid, lipoic acid 50 mg, VB5 50
Mg, 50 mg of heme, 1,2- naphthoquinones, 50 mg, water surplus.
2. charcoal preparation according to claim 1, it is characterised in that：The composition of every liter of SL-10 trace element solution is：
HCl 10.0mL, FeCl2·4H2O 1.5g, ZnCl270.0mg MnCl2·4H2O 100.0mg, H3BO36.0mg, CoCl2·
6H2O 196.0mg, CuCl2·2H2O 2.0mg, NiCl2·6H2O 24.0mg, Na2MoO4·2H2O 36.0mg, water surplus.
3. charcoal preparation according to claim 1, it is characterised in that：The composition of every liter of Se/W solution is：NaOH 0.5
G, NaSeO3·5H2O 3 mg, Na2WO4·2H24 mg of O, water surplus.
4. charcoal preparation according to claim 1, it is characterised in that：The preparation method of the Caragana Microphylla biological charcoal is：It will
Caragana microphylla is cut into the branch of 5-10cm long, and in retort, under oxygen free condition, 400-500 DEG C of temperature carbonizes 4-6h, takes out, machinery
It is broken, grinding, you can obtain Caragana Microphylla biological charcoal.
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| CN107929997B (en) * | 2017-10-13 | 2020-06-23 | 浙江大学 | Application of rape stalk biochar in mediating polychlorinated biphenyl anaerobic reduction dechlorination |
| CN107929998B (en) * | 2017-10-13 | 2020-06-30 | 浙江大学 | Biochar preparation for mediating anaerobic reduction dechlorination of polychlorinated biphenyl |
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