CN110540938A - Ordered oriented co-immobilized enzyme membrane reactor and preparation method and application thereof - Google Patents

Ordered oriented co-immobilized enzyme membrane reactor and preparation method and application thereof Download PDF

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CN110540938A
CN110540938A CN201910618133.8A CN201910618133A CN110540938A CN 110540938 A CN110540938 A CN 110540938A CN 201910618133 A CN201910618133 A CN 201910618133A CN 110540938 A CN110540938 A CN 110540938A
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enzyme
membrane
solution
immobilized
membrane reactor
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叶鹏
祝黛莲
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Zhejiang University of Technology ZJUT
Zhejiang Sci Tech University ZSTU
Zhejiang University of Science and Technology ZUST
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Zhejiang University of Technology ZJUT
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Abstract

the invention discloses an orderly oriented co-immobilized enzyme membrane reactor and a preparation method and application thereof, based on a pervaporation membrane component, the pervaporation membrane component comprises a pervaporation membrane compounded with a microporous filter membrane, immobilized enzymes of combined enzymes required by reaction are sequentially fixed on the microporous filter membrane according to an enzyme catalysis cascade reaction sequence, the combined enzymes are any one of a combination of horseradish peroxidase and glucose oxidase, a combination of tyrosinase and alkaline phosphatase, a combination of long-chain alcohol oxidase and omega-transaminase and a combination of alcohol dehydrogenase, cyclohexanone monooxygenase and lipase, and are beneficial to improving the catalytic reaction efficiency, the immobilized enzymes are integrated on one microporous filter membrane, and the transfer efficiency of multi-step catalytic intermediate products is improved; reaction and separation are integrated by arranging the microporous filter membrane, so that the reaction balance is favorably moved to a favorable direction; the reaction condition is mild, and the enzyme immobilization method is simpler.

Description

ordered oriented co-immobilized enzyme membrane reactor and preparation method and application thereof
Technical Field
The invention belongs to the technical field of reactors, and particularly relates to an orderly oriented co-immobilized enzyme membrane reactor, and a preparation method and application thereof.
Background
enzymes act as biocatalysts, increasing the rate of a wide range of diverse chemical reactions occurring in the body. It is an indispensable component for a living body to participate in important chemical reaction processes essential to life, such as DNA replication and transcription, protein synthesis, primary and secondary metabolism, signal transduction, and cellular defense mechanisms. Enzymes are widely used in the fields of biocatalysis, biosensing, biomedical devices, and the like. However, the enzyme has problems of low thermal stability, poor stability of organic solvents, high cost, poor reusability, etc., so that its wide application is greatly hindered.
The enzyme immobilization technique is one of effective methods for overcoming the disadvantages of free enzymes, and shows a very good effect in improving the stability of enzymes under storage and reaction conditions. People usually fix the immobilized enzyme on different macromolecular carriers by using an enzyme immobilization technology and then prepare the immobilized enzyme reactor, so that the time of enzyme catalytic reaction can be shortened, and the reusability of enzyme is improved.
disclosure of Invention
The invention aims to overcome the problem of low conversion efficiency of the conventional immobilized enzyme reactor and provide an orderly oriented co-immobilized enzyme membrane reactor.
The second purpose of the invention is to overcome the problem of complex preparation method of the existing immobilized enzyme reactor and provide the preparation method of the ordered oriented co-immobilized enzyme membrane reactor.
the invention also aims to provide application of the orderly oriented co-immobilized multienzyme reactor.
the technical scheme for solving the technical problems is as follows:
An orderly oriented co-immobilized enzyme membrane reactor is based on a pervaporation membrane assembly, wherein the pervaporation membrane assembly comprises a pervaporation membrane compounded with a microporous filter membrane, immobilized enzymes of combined enzymes required by reaction are sequentially immobilized on the microporous filter membrane according to an enzyme catalysis cascade reaction sequence, and the combined enzymes are any one of a combination of horseradish peroxidase and glucose oxidase, a combination of tyrosinase and alkaline phosphatase, a combination of long-chain alcohol oxidase and omega-transaminase and a combination of alcohol dehydrogenase, cyclohexanone monooxygenase and lipase.
preferably, the organic macromolecular carrier of the immobilized enzyme is made of a metal organic framework Material (MOF).
preferably, the metal organic framework Material (MOF) is ZIF-8 nanoparticles.
preferably, the pervaporation membrane is a polydimethoxysiloxane/polyvinylidene fluoride (PDMS/PVDF) composite membrane, the microfiltration membrane is a PVDF membrane, and the immobilized enzyme is fixed on the microfiltration membrane by a dead-end filtration method.
Preferably, the aperture of the PVDF membrane is 1-3 μm, and the fixation amount of all immobilized enzymes on the PVDF membrane per unit area is 0.1-0.5 mg/cm 2.
Preferably, the permeation flux of the orderly oriented co-immobilized enzyme membrane reactor is 21-22 g.m < -2 > h < -1 >.
A preparation method of an ordered oriented co-immobilized enzyme membrane reactor is characterized by comprising the following steps:
First, an immobilized enzyme solution of a combination enzyme is prepared
step 1: respectively dissolving the combined enzyme in deionized water to obtain a plurality of enzyme solutions;
step 2: mixing the multiple enzyme solutions obtained in the step 1 with a mixed solution respectively, reacting for 30min at room temperature, and standing for 3h to obtain multiple reaction solutions, wherein the mixed solution comprises a zinc nitrate solution and a 2-methylimidazole solution;
And step 3: centrifuging the multiple reaction solutions obtained in the step 2 at 6000rpm for 10min, and respectively collecting multiple white powders;
And 4, step 4: respectively washing the multiple kinds of white powder obtained in the step 3 with deionized water for 3 times, respectively dispersing the white powder in the deionized water, freeze-drying, and collecting multiple kinds of immobilized enzymes in the form of white powder;
and 5: putting the various white powders into the deionized water again respectively, and performing ultrasonic dispersion to obtain an immobilized enzyme solution of the combined enzyme;
Second, dead-end filtration, fixation and compounding
Fixing the immobilized enzyme solution of the combined enzyme obtained in the first step on a microporous filter membrane through dead-end filtration in sequence, washing with deionized water at least twice, compounding the microporous filter membrane on a pervaporation membrane, and fixing the pervaporation membrane in a pervaporation membrane component.
Preferably, in the step 1, the concentration of any enzyme solution is (3.0-5.2) g/L; in the step 2, the volume ratio of the zinc nitrate solution to the 2-methylimidazole solution to any enzyme solution is 2: 20: (1-2); the concentration of the zinc nitrate solution is 0.25-0.36mol/L, and the concentration of the 2-methylimidazole solution is 1.11-1.42 mol/L.
the application of the ordered oriented co-immobilized enzyme membrane reactor is based on the ordered oriented co-immobilized enzyme membrane reactor, wherein a liquid storage tank is also arranged on one side of the ordered oriented co-immobilized enzyme membrane reactor, the enzyme membrane reactor is connected with the liquid storage tank through a feed pipe and a feed liquid pump, and the same side of the enzyme membrane reactor is also connected with the liquid storage tank through a first discharge pipe; and a cold trap, a drying tower, a buffer bottle and a vacuum pump are sequentially arranged on the other side of the enzyme membrane reactor, the enzyme membrane reactor is connected with the cold trap through a second discharge pipe, a pump is arranged between the cold trap and the drying tower, and the initial substrate passes through the enzyme membrane reactor which is orderly and directionally co-fixed to collect the permeate.
Specifically, the air pressure at the downstream of the pervaporation membrane module is adjusted by a pump between the cold trap and the drying tower, so that the air pressure difference is generated between the upstream and the downstream of the pervaporation membrane, and after the air pressure is adjusted to a certain pressure, the pervaporation is started: the raw materials in the liquid storage tank are conveyed to one side of the enzyme membrane reactor through the feed pipe by the feed liquid pump, the raw materials move to the other side of the enzyme membrane reactor under the action of air pressure difference, so that the raw materials are subjected to cascade reaction under the action of various immobilized enzymes on the microfiltration membrane, reaction substrates are converted to form products, the products are enriched on the other side of the pervaporation membrane to form permeate, and if the reaction substrates conveyed to the pervaporation membrane module by the feed liquid pump are excessive, then the permeate is returned to the liquid storage tank through the first discharge pipe, and is conveyed to the cold trap through the discharge pipe under the action of the pump, dry ice is arranged in the cold trap and is used for liquefying the permeate to form permeate, and for the gas which cannot be liquefied in time, the drying tower absorbs water vapor, and a buffer bottle is arranged in front of the vacuum pump in order to prevent the gas of the drying tower from entering the vacuum pump under the action of the vacuum pump.
Compared with the prior art, the invention has the beneficial effects that:
The arrangement sequence of the immobilized enzymes is in accordance with the catalytic sequence of the enzyme cascade reaction, which is beneficial to improving the catalytic reaction efficiency, and the immobilized enzymes are integrated on a microporous filter membrane, so that the transfer efficiency of the multi-step catalytic intermediate product is improved; reaction and separation are integrated by arranging the microporous filter membrane, so that the reaction balance is favorably moved to a favorable direction; the reaction condition is mild, and the enzyme immobilization method is simpler.
drawings
FIG. 1 is a schematic structural diagram of the present invention.
FIG. 2 is a schematic diagram of an apparatus for converting glucose.
the labels in the figure are: 1-pervaporation membrane component, 2-microporous filter membrane, 3-pervaporation membrane, 4-enzyme membrane reactor, 5-cold trap, 6-pump, 7-drying tower, 8-buffer bottle, 9-vacuum pump, 10-liquid storage tank and 11-liquid pump.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1:
A preparation method of an orderly oriented co-immobilized enzyme membrane reactor comprises the following steps:
First, an immobilized enzyme solution of a combination enzyme is prepared
Step 1: respectively dissolving 5mg of horseradish peroxidase (HRP) molecules and 5mg of Glucose Oxidase (GOD) molecules in 1mL of deionized water to obtain horseradish peroxidase solution and glucose oxidase solution;
In step 1 of the present invention, the glucose oxidase molecule contains a prosthetic Flavin Adenine Dinucleotide (FAD).
step 2: respectively mixing the horseradish peroxidase (HRP) solution and the Glucose Oxidase (GOD) solution in the step 1 with a mixed solution, reacting for 30min at room temperature, and standing for 3h to obtain 2 reaction solutions, wherein the mixed solution comprises 2-3mL of 0.31 mol/L zinc nitrate solution and 20-21mL of 1.36 mol/L2-methylimidazole solution;
And step 3: centrifuging the 2 reaction solutions obtained in the step 2 at 6000r/min for 10min, and respectively collecting 2 white powders;
And 4, step 4: respectively washing 2 kinds of white powder for 3 times by using deionized water, then respectively dispersing the white powder in the deionized water for freeze drying, and collecting white powdery HRP/ZIF-8 and GOD/ZIF-8;
And 5: and (3) respectively putting the HRP/ZIF-8 and the GOD/ZIF-8 obtained in the step (4) into 10mL of deionized water again, and performing ultrasonic dispersion to obtain an HRP/ZIF-8 solution and a GOD/ZIF-8 solution.
second, dead-end filtration, fixation and compounding
and (2) sequentially filtering the HRP/ZIF-8 solution and the GOD/ZIF-8 solution obtained in the first step through a dead end to fix the solutions on a PVDF membrane (the diameter of the membrane is 55mm, and the pore diameter is 2 um), washing the PVDF membrane twice by using deionized water, compounding the PVDF membrane on the PDMS/PVDF membrane, and fixing the PDMS/PVDF membrane in a pervaporation membrane component to obtain the ordered oriented co-immobilized enzyme membrane reactor.
In the step 2 of the invention, the zinc nitrate solution and the 2-methylimidazole solution are mixed, so that zinc ions and 2-methylimidazole are coordinated to form ZIF-8.
The dead-end filtration method in the second step of the invention comprises the following steps: the immobilized enzyme solution is placed at the upstream of the membrane, under the pushing of pressure difference, water and the immobilized enzyme smaller than the membrane pores penetrate the microporous membrane, and the immobilized enzyme larger than the membrane pores is intercepted by the microporous membrane, so that the immobilized enzyme is fixed on the microporous membrane, which is a conventional technical means in the field.
the composite of the PVDF film and the PDMS/PVDF film can be as follows: coating a layer of adhesive on the PVDF film, and then gluing the PVDF film with the PDMS/PVDF film or other composite methods, which are conventional technical means in the field; as for how to fix the pervaporation membrane in the pervaporation membrane module, it is a routine technical means in the art, and a person skilled in the art can set the pervaporation membrane according to the actual situation.
The enzyme membrane reactor with ordered orientation co-fixation is used for decomposing glucose, a liquid storage tank is further arranged on one side of the enzyme membrane reactor, the enzyme membrane reactor is connected with the liquid storage tank through a feed pipe and a feed liquid pump, and the enzyme membrane reactor is also connected with the liquid storage tank through a first discharge pipe on the same side; the other side of the enzyme membrane reactor is sequentially provided with a cold trap, a drying tower, a buffer bottle and a vacuum pump, the enzyme membrane reactor is connected with the cold trap through a second discharge pipe, a pump is arranged between the cold trap and the drying tower, a proper amount of glucose solution with the concentration of 20 mu mol/L is put into a liquid storage tank, the glucose solution in the liquid storage tank is conveyed to the enzyme membrane reactor by a feed liquid pump to serve as a substrate on one side of a pervaporation membrane, the substrate firstly passes through GOD/ZIF-8 under the action of the pump, the glucose is catalyzed by glucose oxidase to generate oxidation reaction under the action of oxidant FAD to generate gluconic acid, the FAD is reduced into FADH2, then the FADH2 transmits hydrogen to oxygen to form hydrogen peroxide, the hydrogen peroxide returns to the FAD, the hydrogen peroxide is catalyzed by the HRP to generate H2O through continuous enzyme catalysis reaction, the H2O obtains a permeate through the other side of the pervaporation membrane, and enters the cold trap under the action, and dry ice is arranged in the cold trap and is used for liquefying the permeate and collecting the permeate.
The reaction process is as follows:
The reaction was continued for 2h at an osmotic flux of 21.05 g.m-2. h-1 with a converted glucose amount of 2.67. mu. mol.
Example 2:
a preparation method of an orderly oriented co-immobilized enzyme membrane reactor comprises the following steps:
First, an immobilized enzyme solution of a combination enzyme is prepared
Step 1: dissolving 4.5mg of Tyrosinase (TYR) molecules and 4.5mg of alkaline phosphatase (AKP) molecules in 1.2mL of deionized water to obtain a tyrosinase solution and an alkaline phosphatase solution, respectively;
Step 2: respectively mixing the tyrosinase solution and the alkaline phosphatase solution obtained in the step 1 with a mixed solution, reacting at room temperature for 30min, and standing for 3h to obtain 2 reaction solutions, wherein the mixed solution comprises 2-3mL of 0.26 mol/L zinc nitrate solution and 20-21mL of 1.13 mol/L2-methylimidazole solution;
and step 3: centrifuging the 2 reaction solutions obtained in the step 2 at 6000r/min for 10min, and respectively collecting 2 white powders;
And 4, step 4: respectively washing 2 kinds of white powder with deionized water for 3 times, respectively dispersing the white powder in the deionized water for freeze drying, and collecting white powdery TYR/ZIF-8 and AKP/ZIF-8;
and 5: and respectively putting TYR/ZIF-8 and AKP/ZIF-8 into the deionized water again, and performing ultrasonic dispersion to obtain TYR/ZIF-8 solution and AKP/ZIF-8 solution.
Second, dead-end filtration, fixation and compounding
and (2) sequentially filtering the TYR/ZIF-8 solution and the AKP/ZIF-8 solution obtained in the first step through a dead end to fix the solutions on a PVDF membrane (the diameter of the membrane is 55mm, and the pore diameter of the membrane is 3 um), washing the PVDF membrane twice by using deionized water, compounding the PVDF membrane on the PDMS/PVDF membrane, and fixing the PDMS/PVDF membrane in a pervaporation membrane component to obtain the ordered oriented co-immobilized enzyme membrane reactor.
The device for decomposing o-phospho-L-tyrosine by using the enzyme membrane reactor is the same as that in the embodiment 1, and the glucose solution in the storage tank is changed into the o-phospho-L-tyrosine solution, and the reaction process is as follows:
the reaction was continued for 2h at a permeation flux of 21.05 gm-2. h-1, converting the amount of o-phospho-L-tyrosine to 3.03. mu. mol.
Example 3:
a preparation method of an orderly oriented co-immobilized enzyme membrane reactor comprises the following steps:
First, an immobilized enzyme solution of a combination enzyme is prepared
step 1: respectively dissolving 5.2mg of long-chain alcohol oxidase (LCAO) molecules and 5.2mg of omega-transaminase (omega-TA) molecules in 1.5mL of deionized water to obtain long-chain alcohol oxidase (LCAO) solution and omega-transaminase (omega-TA) solution;
Step 2: respectively mixing the long-chain alcohol oxidase solution and the omega-transaminase solution obtained in the step 1 with a mixed solution, reacting for 30min at room temperature, and standing for 3h to obtain 2 reaction solutions, wherein the mixed solution comprises 2-3mL of zinc nitrate solution with the concentration of 0.26 mol/L and 20-21mL of 2-methylimidazole solution with the concentration of 1.13 mol/L;
And step 3: centrifuging the 2 reaction solutions obtained in the step 2 at 6000rpm for 10min, and respectively collecting 2 white powders;
and 4, step 4: respectively washing 2 kinds of white powder for 3 times by using deionized water, then respectively dispersing the white powder in the deionized water for freeze drying, and collecting white powdery LCAO/ZIF-8 and omega-TA/ZIF-8;
And 5: and (4) respectively putting the LCAO/ZIF-8 and the omega-TA/ZIF-8 in the deionized water again, and performing ultrasonic dispersion to obtain an LCAO/ZIF-8 solution and an omega-TA/ZIF-8 solution.
second, dead-end filtration, fixation and compounding
And sequentially filtering and fixing the LCAO/ZIF-8 solution and the omega-TA/ZIF-8 solution obtained in the first step on a PVDF membrane (the diameter of the membrane is 55mm, and the pore diameter is 2 um) through a dead end, washing the membrane twice by using deionized water, then compounding the PVDF membrane on the PDMS/PVDF membrane, and finally fixing the PDMS/PVDF membrane in a pervaporation membrane component to obtain the ordered oriented co-immobilized enzyme membrane reactor.
the device for decomposing the long-chain primary aliphatic alcohol by using the enzyme membrane reactor is the same as that in the embodiment 1, the glucose solution in the storage tank is changed into the long-chain primary aliphatic alcohol solution, and the reaction process is as follows:
the reaction is continued for 2 hours under the condition that the permeation flux is 21.05 g.m < -2 > h < -1 >, and the amount of the converted long-chain primary aliphatic alcohol is 4.03 mu mol.
Example 4:
A preparation method of an orderly oriented co-immobilized enzyme membrane reactor comprises the following steps:
first, an immobilized enzyme solution of a combination enzyme is prepared
Step 1: respectively dissolving 5mg of Alcohol Dehydrogenase (ADH) molecules, 5mg of cyclohexanone monooxygenase (CHMO) molecules and 5mg of lipase (CAL-A) molecules in 1mL of deionized water to obtain an alcohol dehydrogenase solution, a cyclohexanone monooxygenase solution and a lipase solution;
Step 2: respectively mixing the ethanol dehydrogenase solution, the cyclohexanone monooxygenase solution and the omega-transaminase solution obtained in the step 1 with a mixed solution, reacting for 30min at room temperature, and standing for 3h to obtain 3 reaction solutions, wherein the mixed solution comprises 2-3mL of 0.31 mol/L zinc nitrate solution and 20-21mL of 1.36 mol/L2-methylimidazole solution;
And step 3: centrifuging the 3 reaction solutions obtained in the step 2 at 6000rpm for 10min, and respectively collecting 3 white powders;
and 4, step 4: and respectively washing 3 kinds of white powder by deionized water for 3 times, respectively dispersing the white powder in the deionized water for freeze drying, and collecting white powder ADH/ZIF-8, CHMO/ZIF-8 and CAL-A/ZIF-8.
And 5: respectively putting ADH/ZIF-8, CHMO/ZIF-8 and CAL-A/ZIF-8 into deionized water again, and performing ultrasonic dispersion to obtain ADH/ZIF-8 solution, CHMO/ZIF-8 solution and CAL-A/ZIF-8 solution;
second, dead-end filtration, fixation and compounding
And successively fixing the ADH/ZIF-8 solution, the CHMO/ZIF-8 solution and the CAL-A/ZIF-8 solution obtained in the first step on a PVDF membrane (the diameter of the membrane is 55mm, and the pore diameter is 2 um) through dead-end filtration, then washing twice with deionized water, then compounding the PVDF membrane on the PDMS/PVDF membrane, and finally fixing the PDMS/PVDF membrane in a pervaporation membrane module to obtain the ordered oriented co-fixed enzyme membrane reactor.
utilize this enzyme membrane reactor to decompose cyclohexanol, the device of decomposition cyclohexanol is the same with embodiment 1, changes the glucose solution in the storage tank into cyclohexanol solution, and the oligomeric reaction of CAL-A alcohol catalysis caprolactone in aqueous phase again after starting substrate cyclohexanol is continuously produced into cyclic caprolactone through ADH and CHMO , and its reaction sequence is as follows:
The reaction was continued for 2h at a permeation flux of 21.05 gm-2. h-1, giving a converted cyclohexanol content of 4.73. mu. mol.

Claims (9)

1. the enzyme membrane reactor is characterized in that the enzyme membrane reactor is based on a pervaporation membrane assembly, the pervaporation membrane assembly comprises a pervaporation membrane compounded with a microporous filter membrane, immobilized enzymes of combined enzymes required by reactions are sequentially fixed on the microporous filter membrane according to an enzyme catalysis cascade reaction sequence, and the combined enzymes are any one of a combination of horseradish peroxidase and glucose oxidase, a combination of tyrosinase and alkaline phosphatase, a combination of long-chain alcohol oxidase and omega-transaminase and a combination of alcohol dehydrogenase, cyclohexanone monooxygenase and lipase.
2. an ordered oriented co-immobilized enzyme membrane reactor according to claim 1 wherein the organic macromolecular support of the immobilized enzyme is made of metal organic framework Material (MOF).
3. The ordered oriented co-immobilized enzyme membrane reactor of claim 2, wherein the metal organic framework Material (MOF) is ZIF-8 nanoparticles.
4. The ordered oriented co-immobilized enzyme membrane reactor of claim 1, wherein the pervaporation membrane is a poly (dimethoxysiloxane)/polyvinylidene fluoride (PDMS/PVDF) composite membrane, the microfiltration membrane is a PVDF membrane, and the immobilized enzyme is immobilized on the microfiltration membrane by dead-end filtration.
5. the ordered oriented co-immobilized enzyme membrane reactor of claim 4, wherein the pore size of the PVDF membrane is 1-3 μm, and the immobilization amount of all immobilized enzymes per unit area of the PVDF membrane is 0.1-0.5 mg/cm 2.
6. The ordered oriented co-immobilized enzyme membrane reactor of claim 1, wherein the enzyme membrane reactor has a permeate flux of 21 to 22 g-m-2-h "1.
7. The process of any of claims 1-6 wherein the process comprises the steps of:
First, an immobilized enzyme solution of a combination enzyme is prepared
Step 1: respectively dissolving the combined enzyme in deionized water to obtain a plurality of enzyme solutions;
step 2: mixing the multiple enzyme solutions obtained in the step 1 with a mixed solution respectively, reacting for 30min at room temperature, and standing for 3h to obtain multiple reaction solutions, wherein the mixed solution comprises a zinc nitrate solution and a 2-methylimidazole solution;
and step 3: centrifuging the multiple reaction solutions obtained in the step 2 at 6000r/min for 10min, and respectively collecting multiple white powders;
And 4, step 4: washing the multiple kinds of white powder obtained in the step 3 with deionized water for 3 times respectively, dispersing the white powder in the deionized water respectively, freeze-drying, and collecting multiple kinds of immobilized enzymes in the form of white powder;
And 5: putting a plurality of immobilized enzymes into the deionized water again respectively, and performing ultrasonic dispersion to obtain an immobilized enzyme solution of the combined enzyme;
second, dead-end filtration, fixation and compounding
Fixing the immobilized enzyme solution of the combined enzyme obtained in the first step on a microporous filter membrane through dead-end filtration in sequence, washing with deionized water at least twice, compounding the microporous filter membrane on a pervaporation membrane, and fixing the pervaporation membrane in a pervaporation membrane component.
8. the method of claim 7, wherein in step 1, the concentration of any enzyme solution is (3.0-5.2) g/L; in the step 2, the volume ratio of the zinc nitrate solution to the 2-methylimidazole solution to any enzyme solution is 2: 20: (1-2); the concentration of the zinc nitrate solution is 0.25-0.36mol/L, and the concentration of the 2-methylimidazole solution is 1.11-1.42 mol/L.
9. The use of the ordered oriented co-immobilized enzyme membrane reactor of any one of claims 1 to 6, wherein based on the ordered oriented co-immobilized enzyme membrane reactor, a liquid storage tank is further provided on one side of the ordered oriented co-immobilized enzyme membrane reactor, the enzyme membrane reactor is connected with the liquid storage tank through a feed pipe and a feed liquid pump, and the enzyme membrane reactor is also connected with the liquid storage tank through a first discharge pipe on the same side; and a cold trap, a drying tower, a buffer bottle and a vacuum pump are sequentially arranged on the other side of the enzyme membrane reactor, the enzyme membrane reactor is connected with the cold trap through a second discharge pipe, a pump is arranged between the cold trap and the drying tower, and the initial substrate passes through the enzyme membrane reactor which is orderly and directionally co-fixed to collect the permeate.
CN201910618133.8A 2019-07-10 2019-07-10 Ordered oriented co-immobilized enzyme membrane reactor and preparation method and application thereof Pending CN110540938A (en)

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Application publication date: 20191206