CN107050732A - A kind of method of utilization white-rot fungi crude enzyme liquid degraded bisphenol AF - Google Patents
A kind of method of utilization white-rot fungi crude enzyme liquid degraded bisphenol AF Download PDFInfo
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- CN107050732A CN107050732A CN201710254666.3A CN201710254666A CN107050732A CN 107050732 A CN107050732 A CN 107050732A CN 201710254666 A CN201710254666 A CN 201710254666A CN 107050732 A CN107050732 A CN 107050732A
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- rot fungi
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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
- A62D2101/22—Organic substances containing halogen
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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 of utilization white-rot fungi crude enzyme liquid degraded bisphenol AF.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 AF methanol solution mixing, set up extracellular enzyme liquid reaction system mixed liquor, through 2 ~ 5d, that is, realize the degraded of bisphenol AF.The inventive method is with low cost, operation is relatively simple, and cost, than relatively low, treating capacity is larger.
Description
Technical field
The invention belongs to environment remediation technical field, more particularly to one kind utilizes microorganism --- and white-rot fungi crude enzyme liquid drops
The method for solving bisphenol AF.
Background technology
White-rot fungi as a kind of microorganism for a variety of difficult degradation compounds that can degrade, bio-pulping, wastewater treatment,
Had a wide range of applications in terms of environment remediation.
Fluoroelastomer is widely used due to its multifunctionality, but generally there are the shortcoming of crosslinking difference.Bisphenol AF（Hexafluoro
Bisphenol-A, C15H10F6O2, BPAF）It is a kind of fluorinated derivatives of bisphenol-A, is widely used as the crosslinking agent and sulphur of fluoroelastomer
Agent, to strengthen the performance of fluoroelastomer.BPAF is also commonly used as multiple polymers such as polyimides, polyamide, polyester, poly- carbon
And the remodeling monomer of other particular polymers, it is usually used in the production of electronic equipment, plastic fiber lamp as monomer.Bisphenol AF and
Other bisphenols are a kind of typical incretion interferent as bisphenol-A, and existing result of study shows, a certain amount of
BPAF is 100% to the terateger rate of zebra fish.Different from bisphenol-A, fluoro organic matter is often more difficult to decompose, and has in the environment
There is higher stability and persistence.As emerging environmental contaminants, BPAF is detected in various environmental matrices, and it leads to
Cross a variety of route of exposure to enter in organism, there is potential risk to ecological and human health.In recent years, BPAF genotoxic potential
The emphasis that environmental area is studied is turned into endocrine disrupting, how to have removed the bisphenol AF in environment and also asked as focus
Topic.So far, less to the degraded report of bisphenol AF, existing optically catalytic TiO 2 degraded bisphenol AF method is to utilize dioxy
Change titanium solution and photocatalytic degradation is carried out to bisphenol AF, but this method degradation time is long, and treating capacity is relatively low, is restricted more, operation
Step is complicated and cost is higher.
Ectoenzyme can be depended on come the principle of degradation of contaminant based on white-rot fungi, it is proposed that one kind uses micro- life
Thing --- white-rot fungi decomposes the new method of bisphenol AF.White-rot fungi growth and breeding is fast, condition of culture gentle, it is more simple to operate
Just.White-rot fungi is cheap and easy to get and green safety, and its ectoenzyme relative amount secreted relatively enriches.Enzymatic reaction has catalysis special
Effect, the features such as green safety, and substrate specificity is extensively, can remove low concentration material, and treated substance concentration range is also more
Extensively, degradation effect can be greatly improved by decomposing bisphenol AF with white-rot fungi, reduce degraded cost, it is in catalytic elimination containing double
There is outstanding advantage on the wastewater processing technology of phenol AF class materials, the mesh for effectively removing environmental contaminants repairing environment can be reached
's.
The content of the invention
The purpose of the present invention be directed to prior art is big to workload present in bisphenol AF degradation process, cost is high,
Treating capacity is low, complex steps the problems such as there is provided a kind of utilization white-rot fungi crude enzyme liquid degrade bisphenol AF method.
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) is obtained moves to 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 times 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 AF
By the MnSO that 0 ~ 2mL concentration is 1 ~ 2mmol/L4Solution, 0 ~ 0.3mL concentration of volume percent are 20% ~ 40% H2O2、1.0
Tartaric acid-sodium tartrate cushioning liquid of ~ 1.5mL the pH value for 4.5, the extracellular thick enzyme of white-rot fungi made from 6 ~ 8mL steps (2)
Liquid and 4 ~ 5 μ L concentration mix for 10g/L bisphenol AF-methanol solution, set up the extracellular enzyme liquid reaction system that cumulative volume is 10mL
Mixed liquor, through 2 ~ 5d, that is, realizes the degraded of bisphenol AF.
The characteristics of the inventive method relies on ectoenzyme degraded environmental contaminants using white-rot fungi, devises a kind of using white
Rotten fungi crude enzyme liquid is degraded the method for bisphenol AF, and selected processing material is only microorganism, growth and breeding is fast, conveniently take,
With low cost, 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 is realized more understand and are apparent to effect, 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, 37 DEG C are placed in, shaken cultivation 5d under the conditions of 130 r/min, OD is controlled 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) is obtained moves to 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
Supernatant after min, centrifugally operated obtains 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 AF
By the MnSO that 1mL concentration is 1mmol/L4Solution, 0.1mL concentration of volume percent are 30% H2O2, 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 AF-methanol solution mixing, sets up extracellular enzyme liquid reaction system mixed liquor, through 3d, that is, realizes the drop of bisphenol AF
Solution.
After tested, the degradation rate of bisphenol AF is 84.4% in the present embodiment.
Claims (1)
- A kind of method of bisphenol AF 1. utilization white-rot fungi crude enzyme liquid is degraded, 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) is obtained moves to 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 times 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 AFBy the MnSO that 0 ~ 2mL concentration is 1 ~ 2mmol/L4Solution, 0 ~ 0.3mL concentration of volume percent are 20% ~ 40% H2O2、1.0 Tartaric acid-sodium tartrate cushioning liquid of ~ 1.5mL the pH value for 4.5, the extracellular thick enzyme of white-rot fungi made from 6 ~ 8mL steps (2) Liquid and 4 ~ 5 μ L concentration mix for 10g/L bisphenol AF-methanol solution, set up the extracellular enzyme liquid reaction system that cumulative volume is 10mL Mixed liquor, through 2 ~ 5d, that is, realizes the degraded of bisphenol AF.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710254666.3A CN107050732A (en) | 2017-04-18 | 2017-04-18 | A kind of method of utilization white-rot fungi crude enzyme liquid degraded bisphenol AF |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| 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 |
-
2017
- 2017-04-18 CN CN201710254666.3A patent/CN107050732A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| 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 |
Non-Patent Citations (4)
| Title |
|---|
| 张飞等: ""木质素过氧化酶对双酚A类化合物的分子动力学模拟"", 《湖北农业科学》 * |
| 暴增海等: "《食用菌栽培学 第1版第1次印刷》", 30 November 1994, 北京教育出版社 * |
| 李胜贤等: "《生物化学》", 30 September 2005, 哈尔滨工业大学出版社 * |
| 秦光军: ""白腐真菌P450降解四溴双酚A的初步研究"", 《硕士学位论文》 * |
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