CN106492396A - A kind of antibiotic bacterium dregs laccase enzyme solution - Google Patents
A kind of antibiotic bacterium dregs laccase enzyme solution Download PDFInfo
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- CN106492396A CN106492396A CN201610850737.1A CN201610850737A CN106492396A CN 106492396 A CN106492396 A CN 106492396A CN 201610850737 A CN201610850737 A CN 201610850737A CN 106492396 A CN106492396 A CN 106492396A
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- antibiotic
- bacterium dregs
- laccase
- antibiotic bacterium
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- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- 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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Abstract
The invention provides antibiotic bacterium dregs are processed, wherein by a kind of method of utilization laccase treatment antibiotic bacterium dregs using laccase：The antibiotic bacterium dregs are sulfur erythromycin bacterium slag；Sulfur erythromycin bacterium slag being processed using laccase, being 40 55 DEG C in temperature, pH is 67, for 4 6h are processed under the conditions of 0.005 0.02g/mL, the potency of sulfur erythromycin bacterium slag reduces by more than 90% to enzyme concentration.The present invention is simple and feasible, and processing cost is low, does not produce secondary pollution, energy-conserving and environment-protective, the antibiotic of most of residual in antibiotic bacterium dregs can be removed, and be easy to the subsequent treatment of antibiotic bacterium dregs, while certain data foundation is provided for ferment treatment antibiotic bacterium dregs technology.
Description
Technical field
The invention belongs to field of Environment Protection, and in particular to a kind of antibiotic bacterium dregs residual antibiotic enzyme solution, more particularly to
One kind is using laccase to antibiotic bacterium dregs residual antibiotic enzyme solution.
Background technology
China is antibiotics big producing country.Substantial amounts of solid waste can be produced in the fermenting and producing of antibiotic,
That is antibiotic bacterium dregs.The main component of antibiotic bacterium dregs is：Produce the microbial mycelial of certain antibiotics, extract antibiotic mistake
The reagent added in journey, the culture medium not being fully used, the degradation product of culture medium, the metabolite produced in sweat, not
Know the antibiotic of somatomedin and a small amount of residual.On the one hand antibiotic bacterium dregs can be caused to air during air storage
Pollution, directly affect the life of inhabitation crowd around；The antibiotic of another aspect bacteria residue residual can enter in soil and
Migrate in soil, so as to increase the drug resistance of microorganism, the health of the remote-effects mankind.Also, in 2008, antibiotic bacterium dregs
It is put into as hazardous waste《National Hazard waste register》.Therefore, find a kind of method of environmental protection and economy to process antibiotic
Bacteria residue is very necessary.
CN105457968A discloses a kind of method for innocent treatment of antibiotic bacterium dregs, and antibiotic bacterium dregs in pH are by which
8～11, inactivate under conditions of 90～100 DEG C of temperature.Bacteria residue is inactivated by this method, and Degradation of Antibiotics therein is lost activity,
The bacteria residue mycelium of inactivation includes cell, flocculation aid and the remaining nutritional labeling of microorganism, you can for organic fertilizer or feeding
Feed additives.But if Degradation of Antibiotics not exclusively, easily causes residual of the antibiotic in organic fertilizer or feed additive,
Connect and be detrimental to health.
CN101380509A discloses a kind of macrolide antibiotic bacterium dregs innocent treatment method, belongs to solid waste
Processing technology field.The method effectively crushes hypha body using adverse current mechanical agitation, then using spiramycin harmless treatment
Microbial inoculum carries out harmless treatment to macrolide antibiotic bacterium dregs, and the antibiotic bacterium dregs that this was processed add as vegetable fertilizer
Plus agent.If but if the interpolation of spiramycin harmless treatment microbial inoculum is improper, easily causing the secondary pollution of antibioticses, and machinery
Stirring has certain cost loss.
Therefore, it is intended that propose one kind can quick, economic, effective method remove residual antibiotic in antibiotic bacterium dregs.
Content of the invention
The purpose of the present invention is essentially consisted in and proposes a kind of new method of antibiotic bacterium Slag treatment, using laccase enzyme to antibiotic
Bacteria residue is processed, and reduces the antibiotic remained in antibiotic bacterium dregs.For solve difficult antibiotic bacterium dregs, high cost,
It is also easy to produce the technical barriers such as secondary pollution, long processing period, complex operation.
The present invention provides a kind of antibiotic bacterium dregs laccase enzyme solution, comprises the following steps：
The erythromycin standard sample of a, compound concentration for 0.001-0.005g/mL；
The antibiotic bacterium dregs suspension of b, compound concentration for 0.01-0.04g/mL, regulation pH are 6-7；
C, in the bacteria residue suspension described in step b plus laccase suspension, be 40-55 DEG C in temperature, the time is 4-6h bars
Antibiotic under part to remaining in antibiotic bacterium dregs is cleared up；
Preferably, the potency of the erythromycin standard sample described in step a is 920-4600U/mL.
Described antibiotic bacterium dregs select sulfur erythromycin bacterium slag, and particular/special requirement is not made in its source.
Preferably, the original titer of bacteria residue suspension described in step b is 73.4-293.7U/mL.
Preferably, in step c, the enzyme concentration of laccase suspension is 0.005-0.02g/mL.
Preferably, the laccase in step c, enzyme activity is 35-140U/mL.
The ratio 1 of laccase suspension and bacteria residue suspension in step c:1.
Wherein, antibiotic is acted on scattering and permeating in Agar Plating, according to the derivation of diffusion law, antibiosis
The logarithm value of plain total amount and antibacterial circle diameter square linear, compare antibiotic standard substance big with the inhibition zone of inspection product
Little, the potency of inspection product antibiotic can be calculated.In the present invention, using the sulfur of erythromycin standard substance and the unknown potency of known potency
The inhibition zone of erythromycin inspection product is compared, then can calculate the potency of sulfur erythromycin bacterium slag.
The present invention compared with prior art, with following remarkable advantage：
The present invention is processed to sulfur erythromycin bacterium slag using laccase, is overcome with sulfur erythromycin bacterium slag as object of study
The bottleneck of specificity ferment treatment specificity substrate.In general, enzyme has strict selectivity to the substrate for being acted on.A kind of enzyme is only
A kind of material, or the material that molecule structure is similar can be acted on, promoting which carries out certain chemical reaction, be produced certain anti-
Product, this selectively acting is answered to be referred to as the specificity of enzyme.Generally enzyme can only be catalyzed a kind of chemical reaction or similar anti-of a class
Should.We conducted substantial amounts of experiment, find the processing method of sulfur erythromycin bacterium slag, but our discoveries for being taken aback, laccase pair
The antibiotic remained in sulfur erythromycin bacterium slag has the excellent effect of clearing up, and the potency of sulfur erythromycin bacterium slag can be made to reduce by 90%
More than.
At present, the method for processing antibiotic bacterium dregs both at home and abroad is mainly, and antibiotic bacterium dregs is carried out at high temperature highly basic inactivation
As organic fertilizer as feed additive or after physical chemistry process after reason, if but chemicals add and improper easily make
Into secondary pollution, and if in bacteria residue, the Degradation of Antibiotics of residual not exclusively easily causes the residual of antibiotic, indirect hazard human body is good for
Health, while the process of high temperature highly basic, physical treatment etc. have certain energy loss, processing cost is higher.Antibiotic bacterium dregs are digested
Method be based on enzyme micro efficient the characteristics of the raw element bacteria residue of antagonism processed, secondary pollution will not be produced, more environmentally friendly, and operate
Simply, reaction time is short, low cost, can remove the antibiotic of most of residual in antibiotic bacterium dregs, be easy to antibiotic bacterium dregs
Subsequent treatment, while providing a kind of new method of ferment treatment antibiotic bacterium dregs.
Specific embodiment
For the technical characterstic of the clearer explanation present invention, the present invention will be carried out in detail by specific embodiment below
Illustrate.
The research of the antibiotic bacterium dregs residual antibiotic laccase zymolysis technique of the present invention, using non-specific enzyme to antibiotic
Bacteria residue is processed, and is comprised the following steps：
The erythromycin standard sample of a, compound concentration for 0.001-0.005g/mL；
The antibiotic bacterium dregs suspension of b, compound concentration for 0.01-0.04g/mL, regulation pH are 6-7；
C, in the bacteria residue suspension plus laccase suspension, be 40-55 DEG C in temperature, the time is to antibiosis under the conditions of 4-6h
The antibiotic remained in plain bacteria residue is cleared up.
Wherein, described antibiotic bacterium dregs select sulfur erythromycin bacterium slag, and original titer is 73.4-293.7U/mL；
Wherein, the enzyme concentration of described laccase suspension is 0.005-0.02g/mL, and enzyme activity is 35-140U/mL
The potency of antibiotic bacterium dregs, the potency of standard substance are determined with cup-plate method；Enzyme activity adopts high effective liquid chromatography for measuring.
Embodiment 1
Erythromycin standard sample of the compound concentration for 0.001g/mL, its potency are 920U/mL；
Sulfur erythromycin bacterium slag suspension of the compound concentration for 0.01g/mL, its original titer are 73.4U/mL, adjust pH and are
6；
Add laccase suspension in the sulfur erythromycin bacterium slag suspension, enzyme concentration is 0.015g/mL, and enzyme activity is 105U/mL；
Temperature be 50 DEG C, the time be 4h under the conditions of in antibiotic bacterium dregs remain antibiotic clear up, after clearing up, sulfur erythromycin
The potency of bacteria residue is 3.67U/mL, and the potency of sulfur erythromycin bacterium slag reduces by 95%.
Embodiment 2
Erythromycin standard sample of the compound concentration for 0.003g/mL, its potency are 2760U/mL；
Sulfur erythromycin bacterium slag suspension of the compound concentration for 0.02g/mL, its original titer are 220.1U/mL, adjust pH and are
7；
Add laccase suspension in the sulfur erythromycin bacterium slag suspension, enzyme concentration is 0.007g/mL, and enzyme activity is 49U/mL；
Temperature be 45 DEG C, the time be 5h under the conditions of in antibiotic bacterium dregs remain antibiotic clear up, after clearing up, sulfur erythromycin
The potency of bacteria residue is 15.4U/mL, and the potency of sulfur erythromycin bacterium slag reduces by 93%.
Embodiment 3
Erythromycin standard sample of the compound concentration for 0.005g/mL, its potency are 4600U/mL；
Sulfur erythromycin bacterium slag suspension of the compound concentration for 0.03g/mL, its original titer are 78.15U/mL, adjust pH and are
7；
Add laccase suspension in the sulfur erythromycin bacterium slag suspension, enzyme concentration is 0.02g/mL, and enzyme activity is 140U/mL；
Temperature be 45 DEG C, the time be 4h under the conditions of in antibiotic bacterium dregs remain antibiotic clear up, after clearing up, sulfur erythromycin
The potency of bacteria residue is 3.9U/mL, and the potency of sulfur erythromycin bacterium slag reduces by 95%.
Embodiment 4
Erythromycin standard sample of the compound concentration for 0.004g/mL, its potency are 3680U/mL；
Sulfur erythromycin bacterium slag suspension of the compound concentration for 0.04g/mL, its original titer are 293.7U/mL, adjust pH and are
6.5；
Add laccase suspension in the sulfur erythromycin bacterium slag suspension, enzyme concentration is 0.015g/mL, and enzyme activity is 105U/mL；
Temperature be 55 DEG C, the time be 6h under the conditions of in antibiotic bacterium dregs remain antibiotic clear up, after clearing up, sulfur erythromycin
The potency of bacteria residue is 5.9U/mL, and the potency of sulfur erythromycin bacterium slag reduces by 98%.
A kind of method of antibiotic bacterium dregs enzymolysis of the present invention, is a kind of new exploration using ferment treatment antibiotic.This
Bright demonstrate the feasibility using ferment treatment antibiotic bacterium dregs method from data, overcome specificity ferment treatment specificity
The bottleneck of bacteria residue, while the method is simple to operate, reaction time is short, low cost, does not produce secondary pollution, with good environmental protection
Benefit.
Claims (7)
1. a kind of antibiotic bacterium dregs laccase enzyme solution, comprises the following steps：
The erythromycin standard sample of a, compound concentration for 0.001-0.005g/mL；
The antibiotic bacterium dregs suspension of b, compound concentration for 0.01-0.04g/mL, regulation pH are 6-7；
C, in the bacteria residue suspension described in step b plus laccase suspension, be 40-55 DEG C in temperature, under the conditions of the time is 4-6h
The antibiotic remained in antibiotic bacterium dregs is cleared up.
2. antibiotic bacterium dregs laccase enzyme solution as claimed in claim 1, it is characterised in that the erythromycin described in step a
The potency of standard sample is 920-4600U/mL.
3. antibiotic bacterium dregs laccase enzyme solution as claimed in claim 1, it is characterised in that described antibiotic bacterium dregs are selected
Sulfur erythromycin bacterium slag.
4. the antibiotic bacterium dregs laccase enzyme solution as described in any one of claims 1 to 3, it is characterised in that described in step b
The original titer of bacteria residue suspension is 73.4-293.7U/mL.
5. the antibiotic bacterium dregs laccase enzyme solution as described in any one of Claims 1 to 4, it is characterised in that laccase in step c
The enzyme concentration of suspension is 0.005-0.02g/mL.
6. the antibiotic bacterium dregs laccase enzyme solution as described in any one of Claims 1 to 5, it is characterised in that the paint in step c
Enzyme, enzyme activity are 35-140U/mL.
7. the antibiotic bacterium dregs laccase enzyme solution as described in any one of claim 1～6, it is characterised in that laccase in step c
The ratio 1 of suspension and bacteria residue suspension:1.
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| CN201610850737.1A CN106492396A (en) | 2016-09-26 | 2016-09-26 | A kind of antibiotic bacterium dregs laccase enzyme solution |
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| CN201610850737.1A CN106492396A (en) | 2016-09-26 | 2016-09-26 | A kind of antibiotic bacterium dregs laccase enzyme solution |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101624605A (en) * | 2008-07-08 | 2010-01-13 | 武汉烁森生态科技有限公司 | Biological method for processing spiramycin bacteria residue |
| CN101979521A (en) * | 2010-09-26 | 2011-02-23 | 吴鹏 | Complex enzyme preparation special for antibiotic dregs |
| CN104328141A (en) * | 2014-10-30 | 2015-02-04 | 宁夏乙征生物工程有限公司 | Method for treating antibiotic residues by enzymic method |
| EP3034186A1 (en) * | 2014-12-16 | 2016-06-22 | Luxembourg Institute of Science and Technology (LIST) | Method of degradation and inactivation of antibiotics in water by immobilized enzymes onto functionalized supports |
-
2016
- 2016-09-26 CN CN201610850737.1A patent/CN106492396A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101624605A (en) * | 2008-07-08 | 2010-01-13 | 武汉烁森生态科技有限公司 | Biological method for processing spiramycin bacteria residue |
| CN101979521A (en) * | 2010-09-26 | 2011-02-23 | 吴鹏 | Complex enzyme preparation special for antibiotic dregs |
| CN104328141A (en) * | 2014-10-30 | 2015-02-04 | 宁夏乙征生物工程有限公司 | Method for treating antibiotic residues by enzymic method |
| EP3034186A1 (en) * | 2014-12-16 | 2016-06-22 | Luxembourg Institute of Science and Technology (LIST) | Method of degradation and inactivation of antibiotics in water by immobilized enzymes onto functionalized supports |
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