CN107412998A - A kind of method for bisphenol S of being degraded using white-rot fungi crude enzyme liquid - Google Patents
A kind of method for bisphenol S of being degraded using white-rot fungi crude enzyme liquid Download PDFInfo
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- CN107412998A CN107412998A CN201710253887.9A CN201710253887A CN107412998A CN 107412998 A CN107412998 A CN 107412998A CN 201710253887 A CN201710253887 A CN 201710253887A CN 107412998 A CN107412998 A CN 107412998A
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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 S.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 S methanol solution mixing, establish extracellular enzyme liquid reaction system mixed liquor, through 2 ~ 5d, that is, realize the degraded of bisphenol S.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 S.
Background technology
Bisphenol S (bisphenol S, BPS), entitled 4, the 4- dihydroxydiphenylsulisomers of chemistry, white, needle-shaped crystals, it is a kind of
Important agricultural chemicals, dyestuff, the intermediate of auxiliary agent.It is various as the product species of Material synthesis using bisphenol S, including the poly- virtue of engineering plastics
Sulfone, bisphenol-s epoxy resin, new-type class H insulating materials polyether sulfone etc., applied available for manufacture laminated product, molding composite, powder
Material flame-retardant etc., there are some researches show, BPS is exposed in environment for a long time, may to the central nervous system of human body and animal,
Immune system, reproductive system and thyroid hormone etc. produce harmful effect.The researcher Wayne of University of California in Los Angeles
Research show that bisphenol S can accelerate embryonic development, destroy the reproductive system of zebra fish.When zebra fish is exposed to the relatively low (phase of concentration
When in the contaminated degree of river) bisphenol-A or bisphenol S when, its brephic physiological function will be sent out in short 25 hours
It is raw to change, there is the presence of bisphenol-A or bisphenol S, embryonic development is faster than normal speed.QIU W etc. research find BPS concentration compared with
In low environment, embryo's normal development of zebra fish is affected, and is embodied in acceleration embryonic development, organism is produced dry
Disturb, cause its premature labor or the probability of precocity to greatly increase, while can also endanger thyroid hormone and female hormone system.Therefore,
It is badly in need of finding a kind of approach for bisphenol S of degrading.Degraded report at present both at home and abroad to bisphenol S is less, and Gao Shengwang etc. passes through synthesis
Magnetic BiOI/Fe3O4And α-the Fe that the bisphenol S that photocatalysis comes in degradation water, Xu Mingjun etc. passes through preparation2O3Ultrathin nanometer piece
And its bisphenol S in visible ray Fenton degradation water, these methods cost in material preparation process is high, complex operation, degradation time
Long, treating capacity is relatively low, is restricted more.The biodegradable green grass or young crops as green new biodegrading process by environmentalist
Look at.But directly carry out degraded complex operation, and easy secondary pollution with bacterial strain, therefore seek a kind of removal efficiency it is high, conveniently,
Safe and free of contamination minimizing technology is imperative.
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.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, main P450 enzymes, the manganese peroxidase of exocytosis for including 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 BPS.
The content of the invention
The purpose of the present invention be directed to prior art is big to workload present in BPS degradation processes, cost is high, processing
A kind of the problems such as measuring low, complex steps, there is provided method for bisphenol S 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 S
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 S-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 S.
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 S, 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, drop
Solution rate is higher.
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 S
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 S-methanol solution mixing, establishes extracellular enzyme liquid reaction system mixed liquor, through 3d, that is, realizes the degraded of bisphenol S.
After tested, the degradation rate of bisphenol S is 94.3% in the present embodiment.
Claims (1)
- A kind of 1. method for bisphenol S 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 SBy 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 S-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 S.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710253887.9A CN107412998A (en) | 2017-04-18 | 2017-04-18 | A kind of method for bisphenol S of being degraded using white-rot fungi crude enzyme liquid |
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| CN201710253887.9A CN107412998A (en) | 2017-04-18 | 2017-04-18 | A kind of method for bisphenol S of being degraded using white-rot fungi crude enzyme liquid |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003334063A (en) * | 2002-05-15 | 2003-11-25 | Kobe Steel Ltd | White rot fungus having high ability to degrade aromatic compound and/or halogenated organic compound |
| JP2006212000A (en) * | 2005-02-07 | 2006-08-17 | Univ Of Fukui | Bacterium for degrading hardly degradable material, and method for cleaning environment by using the same |
| 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 |
| CN101549936A (en) * | 2009-05-04 | 2009-10-07 | 清华大学 | A method using white-rot fungus and carrier to process persistent wastewater |
| 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 CN201710253887.9A patent/CN107412998A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003334063A (en) * | 2002-05-15 | 2003-11-25 | Kobe Steel Ltd | White rot fungus having high ability to degrade aromatic compound and/or halogenated organic compound |
| JP2006212000A (en) * | 2005-02-07 | 2006-08-17 | Univ Of Fukui | Bacterium for degrading hardly degradable material, and method for cleaning environment by using the same |
| 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 |
| CN101549936A (en) * | 2009-05-04 | 2009-10-07 | 清华大学 | A method using white-rot fungus and carrier to process persistent wastewater |
| 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 (2)
| Title |
|---|
| 姚文兵: "《生物技术制药概论》", 31 August 2015, 中国医药科技出版社 * |
| 张甲耀等: "《环境微生物学》", 31 December 2008, 武汉大学出版社 * |
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