CN107050731A - A kind of method of utilization white-rot fungi crude enzyme liquid degrading tetrabromobisphenol A - Google Patents
A kind of method of utilization white-rot fungi crude enzyme liquid degrading tetrabromobisphenol A Download PDFInfo
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- CN107050731A CN107050731A CN201710253886.4A CN201710253886A CN107050731A CN 107050731 A CN107050731 A CN 107050731A CN 201710253886 A CN201710253886 A CN 201710253886A CN 107050731 A CN107050731 A CN 107050731A
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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 of utilization white-rot fungi crude enzyme liquid degrading tetrabromobisphenol A.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 tetrabromobisphenol A methanol solution mixing, set up extracellular enzyme liquid reaction system mixed liquor, through 2 ~ 5d, that is, realize the degraded of tetrabromobisphenol A.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 tetrabromobisphenol A.
Background technology
Tetrabromobisphenol A（Tetrabromobisphenol A, TBBPA）, it is that a kind of bromine class relatively common at present is fire-retardant
Agent, as reactive flame retardant, available for epoxy resin, polyurethane resin etc.；It can be used for polyphenyl second as additive flame retardant
Alkene, SAN resins and ABS resin etc..As common brominated flame retardant, its contamination hazard is obvious.Long Term Contact bromine
Brain and skeleton development can be hindered by changing fire retardant；And when being incinerated processing, the dioxin and furans of bromination can be discharged, the two
It is easily carcinogen.A large amount of uses and its bioconcentration due to tetrabromobisphenol A, in a variety of surrounding mediums and life
Be detected in thing sample, such as air, water body, soil and deposit, some exposed to fire retardant specific crowd tissue,
Blood, breast milk and the general population even arctic all have detected that the presence of tetrabromobisphenol A.Therefore a kind of removal efficiency is sought
It is high, conveniently, safe and free of contamination biodegrading process it is imperative.Conventional biodegrading process has mechanical degradation, photochemistry drop at present
Solution and biodegradable, the undoubtedly most green environmental protection of biological degradation method, thus also most favored by environmental science worker.Qian Yanyuan
The aeration bacteria of tetrabromobisphenol A has been obtained Deng screening and for the Study on degradation to TBBPA；Literary person of outstanding talent's equality, which is then screened, to be obtained
Two plants of anaerobic degrading bacterias are degraded to tetrabromobisphenol A.
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, turns 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.Phanerochaete chrysosporium as white-rot fungi type strain, it is dirty to numerous difficult degradations
Dye thing has significant degradation effect.There are some researches show effect of the Phanerochaete chrysosporium to hard-degraded substance is mostly derived from
The abundant enzyme system of its own secretion, mainly includes P450 enzymes, the manganese peroxidase of exocytosis of emiocytosis（MnP）And wood
Lignin peroxidase（Lip）, enzymatic reaction has the characteristics of efficient green is safe, therefore considers to the thick of white rot fungus secretion
Enzyme liquid is extracted biodegradable to be carried out to tetrabromobisphenol A.
The content of the invention
The purpose of the present invention is to be directed to prior art is big to workload present in tetrabromobisphenol A degradation process, cost
The problems such as height, low treating capacity, complex steps, there is provided a kind of method of utilization white-rot fungi crude enzyme liquid degrading tetrabromobisphenol A.
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 tetrabromobisphenol A
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 tetrabromobisphenol A-methanol solution, set up the extracellular enzyme liquid reactant that cumulative volume is 10mL
It is mixed liquor, through 2 ~ 5d, that is, realizes the degraded of tetrabromobisphenol A.
The characteristics of the inventive method relies on ectoenzyme degraded environmental contaminants using white-rot fungi, devises a kind of using white
The method of rotten fungi crude enzyme liquid degrading tetrabromobisphenol A, selected processing material is only microorganism, and growth and breeding is fast, conveniently take
With, it is with low cost, biology enzyme has efficient catalytic ability.In addition, operation is relatively simple, and cost is than relatively low, treating capacity compared with
Greatly.
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 tetrabromobisphenol A
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 tetrabromobisphenol A-methanol solution mixing, sets up extracellular enzyme liquid reaction system mixed liquor, through 3d, that is, realizes tetrabromobisphenol A
Degraded.
After tested, the degradation rate of tetrabromobisphenol A is 66.1% in the present embodiment.
Claims (1)
1. a kind of method of utilization white-rot fungi crude enzyme liquid degrading tetrabromobisphenol A, it is characterised in that 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 tetrabromobisphenol A
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 tetrabromobisphenol A-methanol solution, set up the extracellular enzyme liquid reactant that cumulative volume is 10mL
It is mixed liquor, through 2 ~ 5d, that is, realizes the degraded of tetrabromobisphenol A.
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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 CN201710253886.4A patent/CN107050731A/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 第1版第1次印刷》", 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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Application publication date: 20170818 |