CN101693873B - Sandwich structuralized enzymatic membrane reactor, production method and application thereof - Google Patents
Sandwich structuralized enzymatic membrane reactor, production method and application thereof Download PDFInfo
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- CN101693873B CN101693873B CN2009100708071A CN200910070807A CN101693873B CN 101693873 B CN101693873 B CN 101693873B CN 2009100708071 A CN2009100708071 A CN 2009100708071A CN 200910070807 A CN200910070807 A CN 200910070807A CN 101693873 B CN101693873 B CN 101693873B
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Abstract
The invention relates to a sandwich structuralized enzymatic membrane reactor and a production method and application thereof. The sandwich structuralized enzymatic membrane reactor is produced by using PES and PEG, or Pluronic F127as raw materials according to mass ratio of 100:10-100:100. The production method comprises the following steps: dissolving the PES and PEG, or Pluronic F127 in DMF and mixing evenly to obtain a casting film liquid; scraping the casting film liquid on flat glass into liquid film and then dipping the liquid film in water, peeling the film, washing with water and dipping; placing the casting film in a film assembly, adding a transglucosidase solution till water in the transglucosidase solution completely penetrates the casting film; laying another compaction filmon transglucosidase, fixing and compacting, i.e, embedding the transglucosidase between the casting film and the compaction film to obtain the sandwich type enzymatic membrane reactor. The invention can effectively prevent enzyme leakage, reduce diffusional limitation effect, and control the substrate transport quantity by controlling pressure and enzyme quantity to match the reaction speed with substrate transport. The invention can be applied to other immobilized enzyme reactions and has wide application range.
Description
Technical field
The present invention relates to the preparation of enzyme mebrane reactor, particularly a kind of sandwich structuralized enzymatic membrane reactor and its production and application.
Background technology
Enzyme mebrane reactor organically combines the high-level efficiency of enzymatic reaction and the selection perviousness of film, can make process intensification.In recent years, enzyme mebrane reactor has obtained increasingly extensive application in fields such as biology, medicine, food, chemical industry, environment.Fixed enzyme membrane reactor has that speed of reaction is fast, selectivity is high, reusable, can eliminate effectively that product suppresses, mass transfer area is big, rate of mass transfer is fast and be easy to serialization, automatic control and advantage such as integrated.The immobilization of enzyme at present has absorption, methods such as covalent attachment, crosslinking and embedding usually.
According to the existence of enzyme, enzyme mebrane reactor can be divided into free state and fixed enzyme membrane reactor two classes.The former enzyme be distributed in equably reactant mutually in, enzymatic reaction is carried out under near the state of intrinsic kinetics, but enzyme is easily sheared inactivation or foam sex change, the remarkably influenced that device performance is polluted by concentration polarization and film.In the fixed enzyme membrane reactor, enzyme is " entrapped " on film by modes such as absorption, crosslinked, embedding, chemical bondings, enzyme packing density height, and performance stability and throughput height, product purity and quality are good, and the refuse amount is few.But enzyme is skewness often, and resistance to mass transfer is also bigger.
Covalent attachment is about to be fixed together by covalent attachment power between enzyme and the upholder, and immobilized enzyme stability improves.The covalent attachment immobilization is improving aspect the thermostability of enzyme, PH stability and the chemical reagent stability even betterly than other process for fixation, and covalent immobilization can also effectively prevent self Degradation of proteolytic enzyme in addition.But covalent immobilization also is most possibly to cause the zymoprotein conformational change, thereby influences the active process for fixation of immobilized enzyme catalysis.
Embedding and microcapsule are different from preceding two kinds of fixing meanss, and the enzyme molecule in its solution is freely, but are fixed on again among gel or the semi-transparent film formed grid structure simultaneously.Can overcome like this absorption fixation method enzyme is come off, prevent enzyme leak into bulk solution mutually in, also overcome deficiencies such as covalent attachment malleable enzyme conformation.But the diffusional limitation that entrapping method and microcapsule method caused is greater than the above two, thereby has reduced enzymatic reaction speed.Microcapsule method still lacks the application of concrete synthetic aspect at present in addition.
Summary of the invention
The object of the present invention is to provide is a kind of sandwich structuralized enzymatic membrane reactor and its production and application, can overcome the deficiencies in the prior art, the design of the invention a kind of " sandwich style " enzyme mebrane reactor, with enzyme molecule double team between ultra-filtration membrane, because the aperture of ultra-filtration membrane has only several nanometers to tens nanometers, can effectively prevent the enzyme leakage phenomenon.Side at film presses simultaneously, is that impellent driving a reaction liquid flows through ultra-filtration membrane with the pressure difference, has reduced diffusion limitation effect, by pressure and enzyme amount control substrate traffic capacity, the transportation of speed of response and substrate is complementary simultaneously.This " sandwich " method also can be applicable in other immobilized enzyme reactions, and the scope of application is extensive.
A kind of sandwich structuralized enzymatic membrane reactor provided by the invention is to be raw material with PES and PEG or Pluronic F127, by mass ratio is preparation in 100: 10～100: 100, concrete steps are: at first PES and PEG or Pluronic F127 are dissolved among the DMF, mix film-casting liquid.Film-casting liquid scraped on sheet glass make liquid film and immerse in the water, this film is peeled off, washing is soaked.Casting film places membrane module to add the complete infiltrate film of water that glucose transglucosidase solution makes enzyme solution, and another piece compacting film is layered on the enzyme, and fixedly compacting is about to enzyme and is embedded between two films, makes " sandwich " formula enzyme mebrane reactor.
The preparation that the invention provides a kind of sandwich structuralized enzymatic membrane reactor that is may further comprise the steps:
1) preparation of PES/PEG and PES/Pluronic F127 film
At first PES and PEG or Pluronic F127 are dissolved among the DMF; the mass ratio of PES and PEG or Pluronic F127 is: between 100: 10～100: 100; guaranteed to mix in 4～6 hours in 50～70 ℃ of following stir abouts, leave standstill about bubble of removing fully in the solution in 4～5 hours then.
2) at room temperature, with film-casting liquid scrape on the sheet glass make uniform liquid film after, immerse in the water immediately, this moment, DMF dissolved in the water, make macromolecular solution be converted into gel-film, this film is stripped down and with deionized water flush away raffinate, be soaked at last in the deionized water about 20～40 hours.
3) immobilization of glucose transglucosidase (transglucosidase)
The film that makes previously is fixed in the membrane module of ultrafiltration cup, be forced under the gauge pressure 0.2MPa pressure water compacting 0.5 hour, again with solution emptying in steel basin and the stationary tank, add rapidly enzyme solution, control pressure is at gauge pressure 0.1MPa, the complete infiltrate film of the water in enzyme solution, and then another block casting film is layered on the enzyme, fixedly compacting is about to enzyme and is embedded between the film, makes " sandwich " formula enzyme mebrane reactor.
The invention provides the efficient production that sandwich structuralized enzymatic membrane reactor is used to realize dextrinosan.
A kind of sandwich structuralized enzymatic membrane reactor provided by the invention can overcome the deficiencies in the prior art, and it between ultra-filtration membrane, because the aperture of ultra-filtration membrane has only several nanometers to tens nanometers, can effectively prevent the enzyme leakage phenomenon with enzyme molecule double team.Side at film presses simultaneously, is that impellent driving a reaction liquid flows through ultra-filtration membrane with the pressure difference, has reduced diffusion limitation effect, by pressure and enzyme amount control substrate traffic capacity, the transportation of speed of response and substrate is complementary simultaneously.This " sandwich " method also can be applied in other immobilized enzyme reactions, and the scope of application is extensive.
Description of drawings
Fig. 1 is the sandwiched type structure enzyme mebrane reactor SEM photo of embodiment 1-3 preparation.
Embodiment
Embodiment 1
Accurately take by weighing 3.60g polyethersulfone (PES) and 1.08g blocked polyethers F127 (Pluronic F127), be dissolved in the N of 15.32g, in the dinethylformamide (DMF), stirred 4 hours down in 60 ℃, leave standstill 5 hours again after.At room temperature, with film-casting liquid scrape on the sheet glass make uniform liquid film after, immerse immediately in the water, behind the gel-film to be formed, this film is stripped down with deionized water flush away raffinate, be soaked at last in the deionized water 20 hours.
The film that makes previously is fixed in the membrane module of ultrafiltration cup, be forced under the gauge pressure 0.15MPa pressure water compacting 0.5 hour, again with solution emptying in steel basin and the stationary tank, add the solution that contains 100mg glucose transglucosidase rapidly, control pressure is at gauge pressure 0.1MPa, and then the complete infiltrate film of water in enzyme solution is layered on another piece film on the enzyme fixedly compacting.Configuration maltose solution 160mg/ml, pH 6.0, adds in the ultrafiltration cup, 50 ℃ of constant temperature, the transformation efficiency that records this enzyme mebrane reactor catalysis maltose is 100%.
Embodiment 2
Accurately take by weighing 3.60g polyethersulfone (PES) and 2.16g blocked polyethers F127 (Pluronic F127), be dissolved in the N of 16.10g, in the dinethylformamide (DMF), stirred 4 hours down in 60 ℃, leave standstill 5 hours again after.At room temperature, with film-casting liquid scrape on the sheet glass make uniform liquid film after, immerse immediately in the water, behind the gel-film to be formed, this film is stripped down with deionized water flush away raffinate, be soaked at last in the deionized water 20 hours.
The film that makes previously is fixed in the membrane module of ultrafiltration cup, be forced under the gauge pressure 0.15MPa pressure water compacting 0.5 hour, again with solution emptying in steel basin and the stationary tank, add the solution that contains the 150mg transfering grape glycosidase rapidly, control pressure is at gauge pressure 0.1MPa, and then the complete infiltrate film of water in enzyme solution is layered on another piece film on the enzyme fixedly compacting.Configuration maltose solution 50mg/ml, pH 5.0, adds in the ultrafiltration cup, 40 ℃ of constant temperature, the transformation efficiency that records this enzyme mebrane reactor catalysis maltose is 32%.
Embodiment 3
Accurately take by weighing 3.60g polyethersulfone (PES) and 0.36g blocked polyethers F127 (Pluronic F127), be dissolved in the N of 16.04g, in the dinethylformamide (DMF), stirred 4 hours down in 60 ℃, leave standstill 5 hours again after.At room temperature, with film-casting liquid scrape on the sheet glass make uniform liquid film after, immerse immediately in the water, behind the gel-film to be formed, this film is stripped down with deionized water flush away raffinate, be soaked at last in the deionized water 20 hours.
The film that makes previously is fixed in the membrane module of ultrafiltration cup, be forced under the gauge pressure 0.15MPa pressure water compacting 0.5 hour, again with solution emptying in steel basin and the stationary tank, add the solution that contains the 200mg transfering grape glycosidase rapidly, control pressure is at gauge pressure 0.1MPa, and then the complete infiltrate film of water in enzyme solution is layered on another piece film on the enzyme fixedly compacting.Configuration maltose solution 150mg/ml, pH 7.0, add in the ultrafiltration cup, and the transformation efficiency that records this enzyme mebrane reactor catalysis maltose is 100%.
Comparative Examples 1
Accurately take by weighing 3.60g polyethersulfone (PES) and 1.08g polyoxyethylene glycol (PEG), be dissolved in the N of 15.32g, in the dinethylformamide (DMF), stirred 4 hours down in 60 ℃, leave standstill 5 hours again after.Under room temperature, with film-casting liquid scrape on the sheet glass make uniform liquid film after, immerse immediately in the water, behind the gel-film to be formed, this film is stripped down with deionized water flush away raffinate, be soaked at last in the deionized water 20 hours.
The film that makes previously is fixed in the membrane module of ultrafiltration cup, be forced under the gauge pressure 0.15MPa water compacting 0.5 hour, again with solution emptying in steel basin and the stationary tank, add the solution that contains 100mg glucose transglucosidase rapidly, control pressure is at gauge pressure 0.1MPa, and then the complete infiltrate film of water in enzyme solution is layered on another piece film on the enzyme fixedly compacting.Configuration maltose solution 160mg/ml, pH 6.0, add in the ultrafiltration cup, and the transformation efficiency that records this enzyme mebrane reactor catalysis maltose is 76%.
Claims (3)
1. the preparation method of a sandwich structuralized enzymatic membrane reactor is characterized in that may further comprise the steps:
1) preparation of PES/PEG and PES/Pluronic F127 film
At first PES and PEG or Pluronic F127 are dissolved among the DMF, the mass ratio of PES and PEG or Pluronic F127 is: between 100: 10～100: 100, guaranteed to mix in 4～6 hours in 50～70 ℃ of following stir abouts, leave standstill about bubble of removing fully in the solution in 4～5 hours then;
2) at room temperature, with film-casting liquid scrape on the sheet glass make uniform liquid film after, immerse in the water immediately, this moment, DMF dissolved in the water, make macromolecular solution be converted into gel-film, this film is stripped down and with deionized water flush away raffinate, be soaked at last in the deionized water 20～40 hours;
3) immobilization of glucose transglucosidase
The film that makes previously is fixed in the membrane module of ultrafiltration cup, be forced under the gauge pressure 0.2MPa pressure water compacting 0.5 hour, again with solution emptying in steel basin and the stationary tank, add rapidly enzyme solution, control pressure is at gauge pressure 0.1MPa, the complete infiltrate film of the water in enzyme solution, and then another block casting film is layered on the enzyme, fixedly compacting is about to enzyme and is embedded between the film, makes sandwich structuralized enzymatic membrane reactor.
2. sandwich structuralized enzymatic membrane reactor that preparation method as claimed in claim 1 makes.
3. sandwich structuralized enzymatic membrane reactor as claimed in claim 2 is characterized in that it is used to realize the preparation of dextrinosan.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4104776A1 (en) * | 1990-08-31 | 1992-03-05 | Inst Molekularbiologie Ak | Sandwich enzyme membrane for enzyme or opto electrodes - consists of of an enzyme layer coated with membrane of inhomogeneous permeability for substrate |
| CN1186115A (en) * | 1996-12-25 | 1998-07-01 | 中国科学院长春应用化学研究所 | Method for biological sensor produced by latex embedding enzyme |
| CN101235351A (en) * | 2008-02-27 | 2008-08-06 | 浙江大学 | Fixed enzyme membrane reactor, preparation thereof and method for producing biological diesel oil by the same |
| CN101265448A (en) * | 2008-04-03 | 2008-09-17 | 浙江大学 | Grease catalysis separation biphasic enzyme-film bioreactor and its preparation and application |
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- 2009-10-15 CN CN2009100708071A patent/CN101693873B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4104776A1 (en) * | 1990-08-31 | 1992-03-05 | Inst Molekularbiologie Ak | Sandwich enzyme membrane for enzyme or opto electrodes - consists of of an enzyme layer coated with membrane of inhomogeneous permeability for substrate |
| CN1186115A (en) * | 1996-12-25 | 1998-07-01 | 中国科学院长春应用化学研究所 | Method for biological sensor produced by latex embedding enzyme |
| CN101235351A (en) * | 2008-02-27 | 2008-08-06 | 浙江大学 | Fixed enzyme membrane reactor, preparation thereof and method for producing biological diesel oil by the same |
| CN101265448A (en) * | 2008-04-03 | 2008-09-17 | 浙江大学 | Grease catalysis separation biphasic enzyme-film bioreactor and its preparation and application |
Non-Patent Citations (1)
| Title |
|---|
| 姜忠义等.酶膜反应器研究进展.《高分子材料科学与工程》.2004,第20卷(第1期),14-17. * |
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