CN110240292A - A method of antibiotic in industrialization laccase and syringaldehyde combined degradation aquiculture sewerage - Google Patents
A method of antibiotic in industrialization laccase and syringaldehyde combined degradation aquiculture sewerage Download PDFInfo
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- CN110240292A CN110240292A CN201910681975.8A CN201910681975A CN110240292A CN 110240292 A CN110240292 A CN 110240292A CN 201910681975 A CN201910681975 A CN 201910681975A CN 110240292 A CN110240292 A CN 110240292A
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- syringaldehyde
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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
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/342—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the enzymes used
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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
- C02F2101/305—Endocrine disruptive agents
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- Environmental & Geological Engineering (AREA)
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
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- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
Abstract
The invention discloses a kind of methods of antibiotic in industrialization laccase and syringaldehyde mediator combined degradation aquiculture sewerage, using the sulfa antibiotics in industrialization 51003 laccase of Novozym and syringaldehyde mediator combined degradation aquiculture sewerage, the following steps are included: adding a certain amount of sulfa antibiotics mother liquor into aquaculture pond sewage, syringaldehyde mediator mother liquor is added again, finally add 51003 laccase of Novozym, adjust suitable pH, oscillating reactions 0.5-6 hours at a certain temperature, complete catalyzing oxidizing degrading reaction, effective degradation can be obtained in sulfa antibiotics in sewage.The present invention is using industrialization laccase as laccase source, and the determination of syringaldehyde mediator concentration has comprehensively considered the ecological effect and degradation effect of mediator, and treatment process is simple, at low cost, pollutants removal rate is high.
Description
Technical field
The invention belongs to biochemical fields, and in particular to it is a kind of using industrialization 51003 laccase of laccase Novozym and
Method of the syringaldehyde as antibiotic in reaction mediator combined degradation aquiculture sewerage.
Background technique
In recent years, the report that antibiotic is detected in water environment constantly draws attention.2015, the first antibiotic row of China
Put the highest attention with the announcement of pollution map by various circles of society.Environment antibiotic residue cause ecological effect problem by
Domestic and international researcher gives more sustained attention.To reduce antibiotic residue to the potential information requirement of environment, especially in recent years at me
State carries out under the background of pollutant discharging license, and the degradation and removal research of the environment contaminant trace species such as antibiotic have become scholars and close
The hot spot cut.
Sulfa antibiotics are common antibacterial and antiphlogistic medicines, are chiefly used in the treatment and prevention of human and animal's disease,
And it is added in animal feed as animal growth promoting agent with asian treatment dosage.Sulfa antibiotics adsorptivity is poor, Neng Gou
Water body stored for extended periods is simultaneously accumulated, and therefore, the discharge that sulfa antibiotics are controlled from source is extremely necessary.
Currently, mainly including both at home and abroad biodegrade and high-level oxidation technology (AOPs) to the degradation technique research of antibiotic
Deng；And removing technical study mainly has traditional removal technology (such as filter, flocculate and settle), absorption, membrane technology and its group
Conjunction method.AOPs technology can efficient degradation multiclass organic pollutant, be the more Degradation of Antibiotics technique of current research.So
And most of AOPs technology single treatment is not thorough in the presence of degradation, higher cost, adds catalyst and oxidant may cause two
Secondary pollution.
To find a kind of cost-effective Degradation of Antibiotics method, in recent years, enzymatic oxidation technology obtains large development,
What is be wherein widely noticed is laccase.Laccase (EC 1.10.3.2) is Japanese scholars Yoshida in point from lac lacquer tree in 1883
A kind of covellite Bovinelactoperoxidase found in secretion.Compared with other oxidizing ferment (such as catalase), in Laccase Catalyzed reaction
O can directly be utilized2As electron acceptor, addition H is not needed2O2, there is the superiority that low energy consumption, high-efficient, environmental-friendly
Energy.Nearly two during the last ten years studies have shown that laccase by be added mediator in the way of can widen significantly laccase apply substrate model
It encloses, has broad application prospects.However, the laccase research reagents of high-purity are mostly used in domestic and foreign literature at present or are painted to producing
Enzyme bacterium is separated and is cultivated, and carries out contaminant degradation research, high-purity in the way of laccase crude extract or immobilization laccase
Laccase it is at high price, and cultivate it is very limited with the yield for separating laccase, which greatly limits the reality of laccase
Application value.
51003 laccase of Novozym is a kind of industrialization laccase of Novozymes Company's production, it is bent by a kind of transgenosis
The laccase preparation that mould prepares through submerged fermentation.Currently, 51003 laccase of Novozym is widely applied in industries such as printing and dyeing, papermaking,
However the application in antibiotics sewage treatment is also rarely reported.
Summary of the invention
It is an object of the invention in 51003 laccase of Novozym and syringaldehyde mediator combined degradation aquiculture sewerage
Antibiotic.Compared with the laccase in other sources, 51003 laccase of Novozym has yield is big, reasonable price, property are stablized etc.
Advantage, these advantages make it be more suitable for actual sewage processing.
Another object of the present invention is to obtain most on the basis of fully considering syringaldehyde itself environment poisonous effect
Excellent degradation parameter solves the practical problem for not considering mediator environmental toxicity in correlative study.
To achieve the goals above, the technical solution of the present invention is as follows: a kind of industrialization laccase combines drop with syringaldehyde mediator
The method for solving antibiotic in aquiculture sewerage, it is characterised in that:
Wherein laccase is 51003 laccase of Novozym, a kind of liquid lacquer of the aspergillus fungi fermenting and producing by transgenosis
Enzyme, laccase activity are greater than 800U/g；It is provided by letter (China) Bioisystech Co., Ltd, Novi, lot number: OMN07027.
Wherein the chemical name of syringaldehyde is syringaldehyde, and No. CAS is 134-96-3, molecule
Amount is 182.17；
Wherein antibiotic is sulfa antibiotics.
A method of antibiotic in industrialization laccase and syringaldehyde mediator combined degradation aquiculture sewerage, feature exist
In the method reaction system by aquiculture sewerage, 51003 laccase group of sulfa antibiotics, syringaldehyde and Novozym
At.Specific reaction method is as follows:
(1) into aquaculture pond sewage, a certain amount of sulfa antibiotics mother liquor is added；
(2) syringaldehyde mediator mother liquor is added again, finally adds 51003 laccase of Novozym；
(3) it adjusts to suitable pH, oscillating reactions 0.5-6 hours at a certain temperature, completes catalyzing oxidizing degrading reaction.
The initial concentration of the sulfa antibiotics is 0.001-0.1g/L.
The syringaldehyde mediator degree need to comprehensively consider its Impacts on ecology and environment and the degradation effect to antibiotic, right according to its
Half effective concentration (the EC of tested material50) determine its ambient concentration upper limit；It is comprehensive to determine in every liter of sewage, the additive amount of syringaldehyde
For 0.05-0.3mmol.
The wherein EC of syringaldehyde50Determine method the following steps are included:
(1) the tested species to the sensitive Fresh Watcr Blue Algae-microcystic aeruginosa of stress as aquatic ecosystem are chosen, pre-
The experimental concentration gradient setting of syringaldehyde is determined on the basis of test；
(2) test (GB/T 21805-2008) method is inhibited using chemicals algal grown, by the verdigris of logarithmic growth phase
Microcystis aeruginosa is exposed under the concentration gradient stress of syringaldehyde, per the biomass for measuring microcystic aeruginosa for 24 hours；
(3) determine syringaldehyde to the EC of microcystic aeruginosa according to microcystic aeruginosa 96h biomass data variation50, syringaldehyde
To the 96hEC of microcystic aeruginosa50For 0.2mM-0.3mM.
Described every liter of sewage additive amount of 51003 laccase of Novozym is 0.5-20ml, i.e., additive amount in the reaction system
Volume ratio is 1/2000-1/50.
The reaction temperature of the reaction system is 5-45 DEG C, pH 2-7, and the revolving speed of oscillation is 100-1000rpm.
The aquiculture sewerage reaches bad V class water water quality.
Innovative point of the invention, mainly: 1, in actual sewage in terms of the removal of antibiotic, with industrialized laccase
It handles, reasonable price is easy to operate, be conducive to practical application；2, the poisonous effect of syringaldehyde has been comprehensively considered, with its half effect
Answer concentration EC50As the actual concentrations SC service ceiling of syringaldehyde, optimal degradation technique parameter is found within this range.
Beneficial effects of the present invention: Laccase Catalyzed oxidation technology degradation of contaminant is with low energy consumption, high-efficient, environmental-friendly
Etc. superior functions, laccase can survive in air, using oxygen as electron acceptor, after reacting, unique reduzate is just
It is water, therefore laccase is substantially a kind of environment-friendly type ferment.The present invention is using industrialized laccase as laccase source, industrialization
Laccase yield is big, property is stablized, and can substantially reduce the consuming cost of laccase in practical applications, be in practical treatment of sewage
Ideal laccase source.Laccase Catalyzed oxidation stain object is assisted as mediator using syringaldehyde, can largely promote pollution
The degradation effect of object.Meanwhile although syringaldehyde is a kind of natural products, to aquatic ecosystem, there are still certain toxicity wind for it
Danger.Therefore, the present invention has comprehensively considered the poisonous effect of syringaldehyde, with its half effect concentration EC50Reality as syringaldehyde is dense
SC service ceiling is spent, finds optimal degradation technique parameter within this range.The study found that under suitable reaction temperature and pH,
Laccase can show preferable bioactivity, to show to the preferable degradation effect of antibiotic.In addition, to actual sewage
In terms of the removal of middle antibiotic, handled with industrialized laccase, reasonable price is easy to operate, is conducive to practical application.With
Into research, industrial laccase is basically used for paper-making fibre industry, treatment of dyeing wastewater, less to be used for other sewage especially antibiosis
The degradation of element.
In general, treatment process of the present invention is simple, at low cost, pollutants removal rate is high.
Specific embodiment
The present invention is such to work and implements, a kind of industrialization laccase and the combined degradation aquaculture of syringaldehyde mediator
A kind of method of antibiotic in sewage, it is characterised in that: wherein laccase is 51003 laccase of Novozym, is song by transgenosis
The activity of the mould liquid laccase for belonging to fungi fermentation production, laccase is greater than 800U/g；Wherein the chemical name of syringaldehyde is 3,5- bis-
Methoxyl group -4- hydroxy benzaldehyde, No. CAS is 134-96-3, molecular weight 182.17；Wherein antibiotic is sulfa antibiotics.
It is characterized in that method and step is as follows:
(1) into aquaculture pond sewage, a certain amount of sulfa antibiotics mother liquor is added；
(2) syringaldehyde mediator mother liquor is added again, finally adds 51003 laccase of Novozym；
(3) it adjusts to suitable pH, oscillating reactions 0.5-6 hours at a certain temperature, completes catalyzing oxidizing degrading reaction.
Below by embodiment, technical solution of the present invention is further described in detail.
Embodiment 1: Degradation of Antibiotics system 100ml successively adds aquiculture sewerage 96,95,94ml, 1g/L antibiotic
Mother liquor respectively adds 1ml, and 10mM syringaldehyde mediator mother liquor adds 1 (0.1mM), 2 (0.2mM), 3ml (0.3mM), Novozym51003 respectively
Laccase respectively adds 2ml, 3 Duplicate Samples of every group of setting.Each system pH=6 is adjusted respectively, is put into temperature control shaken cultivation case, temperature is
25 DEG C, revolving speed 150rpm.It takes 1ml sample to 5ml centrifuge tube respectively in 180min, adds 1ml methanol mixing progress laccase and go out
Processing living is to stop reacting.Most afterwards after 0.22 μm of syringe filters filters, it is transferred to 2ml brown sample injection bottle and carries out remaining antibiosis
The measurement of plain concentration.Syringaldehyde mediator is not added as a control group.Degradation of Antibiotics effect is as follows:
Embodiment 2: Degradation of Antibiotics system 100ml adds aquiculture sewerage 97ml, the addition of 1g/L antibiotic mother liquor
1ml, 10mM syringaldehyde mediator mother liquor add 1.87ml (0.187mM), and 51003 laccase of Novozym adds 0.051ml, and every group sets
Set 3 Duplicate Samples.Reaction system pH=5.5 is adjusted, is put into temperature control shaken cultivation case, temperature is 30.3 DEG C, revolving speed 150rpm.
It takes 1ml sample to 5ml centrifuge tube respectively in 180min, adds 1ml methanol and mix progress laccase inactivation treatment to stop reacting.
Most afterwards after 0.22 μm of syringe filters filters, it is transferred to the measurement that 2ml brown sample injection bottle carries out remaining antibiotic concentration.With not
Add syringaldehyde mediator as a control group.Degradation of Antibiotics effect is as follows:
Embodiment 3: Degradation of Antibiotics system 100ml adds aquiculture sewerage 96.9ml, the addition of 1g/L antibiotic mother liquor
1ml, 10mM syringaldehyde mediator mother liquor add 2ml (0.2mM), 51003 laccase of Novozym add 0.088ml, every group setting 3
Duplicate Samples.Reaction system pH=5.3 is adjusted, is put into temperature control shaken cultivation case, temperature is 31 DEG C, revolving speed 150rpm.?
It takes 1ml sample to 5ml centrifuge tube when 180min respectively, adds 1ml methanol and mix progress laccase inactivation treatment to stop reacting.Most
After filtering by 0.22 μm of syringe filters, it is transferred to the measurement that 2ml brown sample injection bottle carries out remaining antibiotic concentration.To be not added
Syringaldehyde mediator is as a control group.Degradation of Antibiotics effect is as follows:
Embodiment 4: Degradation of Antibiotics system 100ml adds aquiculture sewerage 97.45ml, and 1g/L antibiotic mother liquor adds
Add 1ml, 10mM syringaldehyde mediator mother liquor adds 1.5ml (0.15mM), and 51003 laccase of Novozym adds 0.05ml, and every group sets
Set 3 Duplicate Samples.Reaction system pH=5.3 is adjusted, is respectively put into temperature control shaken cultivation case, 15,30,45 DEG C of temperature, is turned
Fast 150rpm.1ml sample is taken respectively in 180min to 5ml centrifuge tube, add 1ml methanol mix carry out laccase inactivation treatment with
Stop reaction.Most afterwards after 0.22 μm of syringe filters filters, it is transferred to 2ml brown sample injection bottle and carries out remaining antibiotic concentration
Measurement.Syringaldehyde mediator is not added as a control group.Degradation of Antibiotics effect is as follows:
Embodiment 5: Degradation of Antibiotics system 100ml adds aquiculture sewerage 97.45ml, and 1g/L antibiotic mother liquor adds
Add 1ml, 10Mm syringaldehyde mediator mother liquor adds 1.5ml (0.15mM), and 51003 laccase of Novozym adds 0.05ml, and every group sets
Set 3 Duplicate Samples.Reaction system pH=2 and 7 is adjusted respectively, is put into temperature control shaken cultivation case, and temperature is 30 DEG C, revolving speed
150rpm.It takes 1ml sample to 5ml centrifuge tube respectively in 180min, adds 1ml methanol and mix progress laccase inactivation treatment to stop
Only react.Most afterwards after 0.22 μm of syringe filters filters, it is transferred to the survey that 2ml brown sample injection bottle carries out remaining antibiotic concentration
It is fixed.Syringaldehyde mediator is not added as a control group.Degradation of Antibiotics effect is as follows:
It is important to note that above embodiments are merely to illustrate, according to the explanation of this paper, in the scope of the present invention
The interior modifications and variations that various parameters are made to the present invention, and other type antibiotic that the method described in the invention carries out
And the degradation of other type effluent sewages, also it is included in the scope of the present invention.
Claims (7)
1. the method for antibiotic, feature exist in a kind of industrialization laccase and syringaldehyde mediator combined degradation aquiculture sewerage
In:
Wherein laccase is 51003 laccase of Novozym, is a kind of liquid laccase of aspergillus fungi fermenting and producing by transgenosis,
The activity of laccase is greater than 800U/g；
Wherein the chemical name of syringaldehyde is syringaldehyde, and No. CAS is 134-96-3, and molecular weight is
182.17；
Wherein antibiotic is sulfa antibiotics.
2. antibiosis in a kind of industrialization laccase according to claim 1 and syringaldehyde mediator combined degradation aquiculture sewerage
The method of element, it is characterised in that steps are as follows for reaction system:
(1) into aquaculture pond sewage, a certain amount of sulfa antibiotics mother liquor is added；
(2) a certain amount of syringaldehyde mediator mother liquor is added again, finally adds a certain amount of 51003 laccase of Novozym；
(3) it adjusts to suitable pH, oscillating reactions 0.5-6 hours at a certain temperature, completes catalyzing oxidizing degrading reaction.
3. antibiosis in a kind of industrialization laccase according to claim 2 and syringaldehyde mediator combined degradation aquiculture sewerage
The method of element, it is characterised in that: the initial concentration of the sulfa antibiotics is 0.001-0.1g/L.
4. antibiosis in a kind of industrialization laccase according to claim 2 and syringaldehyde mediator combined degradation aquiculture sewerage
The method of element, it is characterised in that: the syringaldehyde mediator concentration need to comprehensively consider its Impacts on ecology and environment and the drop to antibiotic
Effect is solved, according to its half effective concentration, that is, EC to tested material50Determine its ambient concentration upper limit；It is finally comprehensive to determine every liter of dirt
In water, the additive amount of syringaldehyde mediator is 0.05-0.3mmol.
5. antibiosis in a kind of industrialization laccase according to claim 4 and syringaldehyde mediator combined degradation aquiculture sewerage
The method of element, it is characterised in that: the wherein EC of syringaldehyde50Determine method the following steps are included:
(1) the tested species to the sensitive Fresh Watcr Blue Algae-microcystic aeruginosa of stress as aquatic ecosystem are chosen, in trial test
On the basis of determine syringaldehyde experimental concentration gradient setting；
(2) test method is inhibited using chemicals algal grown, the microcystic aeruginosa of logarithmic growth phase is exposed to syringaldehyde
Under concentration gradient stress, per the biomass for measuring microcystic aeruginosa for 24 hours；
(3) determine syringaldehyde to the EC of microcystic aeruginosa according to microcystic aeruginosa 96h biomass data variation50, syringaldehyde is to copper
The 96hEC of green Microcystis aeruginosa50For 0.2-0.3mM.
6. antibiosis in a kind of industrialization laccase according to claim 2 and syringaldehyde mediator combined degradation aquiculture sewerage
The method of element, it is characterised in that: described every liter of sewage additive amount of 51003 laccase of Novozym is 0.5-20ml, and conversion is being reacted
Additive amount volume ratio in system is 1/2000-1/50.
7. antibiosis in a kind of industrialization laccase according to claim 2 and syringaldehyde mediator combined degradation aquiculture sewerage
The method of element, it is characterised in that: the reaction temperature of the reaction system is 5-45 DEG C, pH 2-7, and the revolving speed of oscillation is 100-
1000rpm。
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Cited By (2)
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| CN115057540A (en) * | 2022-07-06 | 2022-09-16 | 吉林大学 | Phenol sewage treatment method based on nano enzyme UMP-Cu with laccase activity |
| CN115872486A (en) * | 2022-12-28 | 2023-03-31 | 江西省水利科学院 | Device and method for removing trace antibiotics in sewage through enrichment and oxidation |
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Cited By (3)
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| CN115057540A (en) * | 2022-07-06 | 2022-09-16 | 吉林大学 | Phenol sewage treatment method based on nano enzyme UMP-Cu with laccase activity |
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