CN107412999A - A kind of method for bisphenol-c of being degraded using white-rot fungi crude enzyme liquid - Google Patents
A kind of method for bisphenol-c of being degraded using white-rot fungi crude enzyme liquid Download PDFInfo
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- CN107412999A CN107412999A CN201710254659.3A CN201710254659A CN107412999A CN 107412999 A CN107412999 A CN 107412999A CN 201710254659 A CN201710254659 A CN 201710254659A CN 107412999 A CN107412999 A CN 107412999A
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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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- 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
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/20—Organic substances
- A62D2101/28—Organic substances containing oxygen, sulfur, selenium or tellurium, i.e. chalcogen
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Abstract
The invention discloses a kind of method using white-rot fungi crude enzyme liquid degraded bisphenol-c.White-Rot Fungi freeze-dried powder is in potato dextrose agar slant culture medium, the strain spore turned out after activation is stand-by, the strain spore after stand-by activation is swept away with sterilized water during inoculation, it is seeded to liquid potato dextrose culture-medium, shaken cultivation, obtain being inoculated with the culture medium of white-rot fungi, move to Centrifuge Cup, centrifugation obtains supernatant and mycelium, remove lower floor's mycelium, by supernatant centrifugally operated 1 ~ 2 time, the supernatant after centrifugally operated obtains the extracellular crude enzyme liquid of white-rot fungi through 0.45 μm of membrane filtration, by MnSO4Solution, H2O2, tartaric acid sodium tartrate cushioning liquid, the extracellular crude enzyme liquid of white-rot fungi and bisphenol-c methanol solution mixing, establish extracellular enzyme liquid reaction system mixed liquor, through 2 ~ 5d, that is, realize the degraded of bisphenol-c.The inventive method cost is cheap, operation is relatively simple, and cost, than relatively low, treating capacity is larger.
Description
Technical field
It is more particularly to a kind of to utilize microorganism the invention belongs to environment remediation technical field --- white-rot fungi crude enzyme liquid drops
The method for solving bisphenol-c.
Background technology
Bisphenol-c（Bisphenol C, BPC）, as environment incretion interferent（endocrine disrupting
Chemicals, EDCs）One of Typical Representative, have the function that, similar in appearance to estrogen, to take in vivo with regard to human body can be destroyed on a small quantity
Internal system, cause infertility, monster etc., its bio-toxicity and endocrine disrupting activity equivalent to bisphenol-A, and they
As emerging chemical materials, there is wide application prospect.After bisphenol-c compound enters human body, fat can be dissolved in and then in body
Interior enrichment, it is difficult to exclude in vitro, there are some researches show the presence that bisphenol-c is found in human serum.Therefore, it is badly in need of finding a kind of drop
Solve the approach of bisphenol-c.The degraded both at home and abroad to bisphenol-c is rarely reported at present, how to reduce this harm, which type of side passed through
Method is come this pollutant of degrading, or it is a section anxious to be resolved to be translated into other compounds pollution-free, that harm is small
Learn one of problem.
White-rot fungi is the general designation for the filamentous fungi that can cause wooden white rot, and majority is sought in timber or soil
Saprogenesis, turn into environment pollution control neck because of its degradation powerful to numerous pollutants especially hardly degraded organic substance
One of domain research object of greatest concern.Type strain of the Phanerochaete chrysosporium as white-rot fungi, it is dirty to numerous difficult degradations
Dye thing has significant degradation effect.But in actual production life, directly it is metabolized difficult degradation compound cost height, behaviour with bacterial strain
Make complexity, easily cause secondary pollution, the defects of certain be present.There are some researches show Phanerochaete chrysosporium is to difficult degradation thing
The effect of matter is mostly derived from the abundant enzyme system of its own secretion, the main P450 enzymes including emiocytosis, the manganese mistake of exocytosis
Oxide enzyme（MnP）And lignin peroxidase（Lip）, enzymatic reaction has the characteristics of efficient green is safe, therefore considers
The crude enzyme liquid of white rot fungus secretion is extracted to carry out biodegradation to bisphenol-c.
The content of the invention
The purpose of the present invention be directed to prior art is big to workload present in BPC degradation processes, cost is high, processing
A kind of the problems such as measuring low, complex steps, there is provided method for bisphenol-c of being degraded using white-rot fungi crude enzyme liquid.
Concretely comprise the following steps：
(1) culture of white-rot fungi
White-Rot Fungi freeze-dried powder is in potato dextrose agar slant culture medium, after 35 ~ 39 DEG C are cultivated 3 ~ 7d, in sterile bar
It is stand-by as the strain spore after activation in 2 ~ 6 DEG C of preservations under part, with sterilized water by the strain spore after stand-by activation during inoculation
Son is swept away, and is seeded to liquid potato dextrose culture-medium, be placed in 34 ~ 39 DEG C, shaken cultivation 4 under the conditions of 110 ~ 140 r/min ~
6d, it is 1.5 that OD values are controlled during inoculation, obtains being inoculated with the culture medium of white-rot fungi.
(2) preparation of the extracellular crude enzyme liquid of white-rot fungi
The culture medium for being inoculated with white-rot fungi that step (1) obtains is moved into Centrifuge Cup, with 2500 ~ 3500 r/min speed from
The min of the heart 5 ~ 15, obtains supernatant and mycelium, removes lower floor's mycelium, by supernatant at 3 ~ 4 DEG C with 8000 ~ 9000 r/
The min min of centrifugation 6 ~ 20, the centrifugally operated number of supernatant is 1 ~ 2 time, and the supernatant after centrifugally operated is through 0.45 μm
Membrane filtration obtains the extracellular crude enzyme liquid of white-rot fungi, and 0 ~ 6 DEG C saves backup.
(3) degraded of bisphenol-c
By the MnSO that 0 ~ 2mL concentration is 1 ~ 2mmol/L4Solution, the H that 0 ~ 0.3mL concentration of volume percent is 20% ~ 40%2O2、1.0
Tartaric acid-sodium tartrate cushioning liquid that ~ 1.5mL pH value is 4.5, the extracellular thick enzyme of white-rot fungi made from 6 ~ 8mL steps (2)
Liquid and bisphenol-c-methanol solution mixing that 4 ~ 5 μ L concentration are 10g/L, establish the extracellular enzyme liquid reaction system that cumulative volume is 10mL and mix
Liquid is closed, through 2 ~ 5d, that is, realizes the degraded of bisphenol-c.
The characteristics of the inventive method relies on ectoenzyme degraded environmental contaminants using white-rot fungi, devise a kind of using white
The method of rotten fungi crude enzyme liquid degraded bisphenol-c, selected processing material are only microorganism, growth and breeding is fast, conveniently take, into
This is cheap, and biology enzyme has efficient catalytic ability.In addition, operation is relatively simple, and cost, than relatively low, treating capacity is larger.
Embodiment
Embodiment：
In order that technological means, reached purpose that the present invention realizes more understand and are apparent to acting on, with reference to specific
The present invention is further elaborated for embodiment, and listed embodiment is only that the explanation present invention without limiting the present invention.
(1) culture of white-rot fungi
White-Rot Fungi freeze-dried powder is in potato dextrose agar slant culture medium, after 37 DEG C are cultivated 5d, aseptically
It is stand-by as the strain spore after activation in 4 DEG C of preservations, the strain spore after stand-by activation is rushed with sterilized water during inoculation
Under, liquid potato dextrose culture-medium is seeded to, is placed in 37 DEG C, shaken cultivation 5d under the conditions of 130 r/min, controls OD during inoculation
It is worth for 1.5, obtains being inoculated with the culture medium of white-rot fungi.
(2) preparation of the extracellular crude enzyme liquid of white-rot fungi
The culture medium for being inoculated with white-rot fungi that step (1) obtains is moved into Centrifuge Cup, with 3000 r/min centrifugation 10
Min, obtains supernatant and mycelium, removes lower floor's mycelium, by supernatant at 4 DEG C with 9000 r/min centrifugation 10
Min, the supernatant after centrifugally operated obtain the extracellular crude enzyme liquid of white-rot fungi through 0.45 μm of membrane filtration, and 4 DEG C save backup.
(3) degraded of bisphenol-c
By the MnSO that 1mL concentration is 1mmol/L4Solution, the H that 0.1mL concentration of volume percent is 30%2O2, 1.4mL pH value be
The extracellular crude enzyme liquid of white-rot fungi made from 4.5 tartaric acid-sodium tartrate cushioning liquid, 7.5mL steps (2) and 5 μ L concentration are
10g/L bisphenol-c-methanol solution mixing, establishes extracellular enzyme liquid reaction system mixed liquor, through 3d, that is, realizes the degraded of bisphenol-c.
After tested, the degradation rate of bisphenol-c is 52.6% in the present embodiment.
Claims (1)
- A kind of 1. method for bisphenol-c of being degraded using white-rot fungi crude enzyme liquid, it is characterised in that concretely comprise the following steps：(1) culture of white-rot fungiWhite-Rot Fungi freeze-dried powder is in potato dextrose agar slant culture medium, after 35 ~ 39 DEG C are cultivated 3 ~ 7d, in sterile bar It is stand-by as the strain spore after activation in 2 ~ 6 DEG C of preservations under part, with sterilized water by the strain spore after stand-by activation during inoculation Son is swept away, and is seeded to liquid potato dextrose culture-medium, be placed in 34 ~ 39 DEG C, shaken cultivation 4 under the conditions of 110 ~ 140 r/min ~ 6d, it is 1.5 that OD values are controlled during inoculation, obtains being inoculated with the culture medium of white-rot fungi；(2) preparation of the extracellular crude enzyme liquid of white-rot fungiThe culture medium for being inoculated with white-rot fungi that step (1) obtains is moved into Centrifuge Cup, with 2500 ~ 3500 r/min speed from The min of the heart 5 ~ 15, obtains supernatant and mycelium, removes lower floor's mycelium, by supernatant at 3 ~ 4 DEG C with 8000 ~ 9000 r/ The min min of centrifugation 6 ~ 20, the centrifugally operated number of supernatant is 1 ~ 2 time, and the supernatant after centrifugally operated is through 0.45 μm Membrane filtration obtains the extracellular crude enzyme liquid of white-rot fungi, and 0 ~ 6 DEG C saves backup；(3) degraded of bisphenol-cBy the MnSO that 0 ~ 2mL concentration is 1 ~ 2mmol/L4Solution, the H that 0 ~ 0.3mL concentration of volume percent is 20% ~ 40%2O2、1.0 Tartaric acid-sodium tartrate cushioning liquid that ~ 1.5mL pH value is 4.5, the extracellular thick enzyme of white-rot fungi made from 6 ~ 8mL steps (2) Liquid and bisphenol-c-methanol solution mixing that 4 ~ 5 μ L concentration are 10g/L, establish the extracellular enzyme liquid reaction system that cumulative volume is 10mL and mix Liquid is closed, through 2 ~ 5d, that is, realizes the degraded of bisphenol-c.
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| JP2002018480A (en) * | 2000-07-10 | 2002-01-22 | Idemitsu Kosan Co Ltd | Method for cleaning water contaminated by refractory material |
| CN1390161A (en) * | 1999-11-11 | 2003-01-08 | 出光兴产业株式会社 | Method of degrading hardly degradable harmful material |
| CN101469312A (en) * | 2007-12-24 | 2009-07-01 | 河南仰韶生化工程有限公司 | White rot fungi, breeding method and use thereof |
| CN103964567A (en) * | 2014-05-22 | 2014-08-06 | 南京大学 | Method for removing pharmaceutical and personal care product (PPCP) pollutants by using lignin peroxidase preparation |
| CN105233458A (en) * | 2015-10-19 | 2016-01-13 | 桂林理工大学 | Method for degrading bisphenol A through white-rot fungus crude enzyme |
| CN106006994A (en) * | 2016-05-20 | 2016-10-12 | 佛山市聚成生化技术研发有限公司 | Method for degrading bisphenol A by virtue of crude enzyme liquid of lentinula edodes |
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| CN1390161A (en) * | 1999-11-11 | 2003-01-08 | 出光兴产业株式会社 | Method of degrading hardly degradable harmful material |
| JP2002018480A (en) * | 2000-07-10 | 2002-01-22 | Idemitsu Kosan Co Ltd | Method for cleaning water contaminated by refractory material |
| CN101469312A (en) * | 2007-12-24 | 2009-07-01 | 河南仰韶生化工程有限公司 | White rot fungi, breeding method and use thereof |
| CN103964567A (en) * | 2014-05-22 | 2014-08-06 | 南京大学 | Method for removing pharmaceutical and personal care product (PPCP) pollutants by using lignin peroxidase preparation |
| CN105233458A (en) * | 2015-10-19 | 2016-01-13 | 桂林理工大学 | Method for degrading bisphenol A through white-rot fungus crude enzyme |
| CN106006994A (en) * | 2016-05-20 | 2016-10-12 | 佛山市聚成生化技术研发有限公司 | Method for degrading bisphenol A by virtue of crude enzyme liquid of lentinula edodes |
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