CN107418949A - A kind of carrier material fixed for cellulase and preparation method thereof - Google Patents

A kind of carrier material fixed for cellulase and preparation method thereof Download PDF

Info

Publication number
CN107418949A
CN107418949A CN201710639092.1A CN201710639092A CN107418949A CN 107418949 A CN107418949 A CN 107418949A CN 201710639092 A CN201710639092 A CN 201710639092A CN 107418949 A CN107418949 A CN 107418949A
Authority
CN
China
Prior art keywords
cellulase
carrier material
preparation
polyacrylonitrile
complex microsphere
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201710639092.1A
Other languages
Chinese (zh)
Inventor
冯海燕
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Suzhou Kanbon Biological Technology Co Ltd
Original Assignee
Suzhou Kanbon Biological Technology Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Suzhou Kanbon Biological Technology Co Ltd filed Critical Suzhou Kanbon Biological Technology Co Ltd
Priority to CN201710639092.1A priority Critical patent/CN107418949A/en
Publication of CN107418949A publication Critical patent/CN107418949A/en
Pending legal-status Critical Current

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/14Enzymes or microbial cells immobilised on or in an inorganic carrier
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01JCHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J13/00Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
    • B01J13/02Making microcapsules or microballoons
    • B01J13/04Making microcapsules or microballoons by physical processes, e.g. drying, spraying
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01JCHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J13/00Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
    • B01J13/02Making microcapsules or microballoons
    • B01J13/04Making microcapsules or microballoons by physical processes, e.g. drying, spraying
    • B01J13/043Drying and spraying
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01JCHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J13/00Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
    • B01J13/02Making microcapsules or microballoons
    • B01J13/06Making microcapsules or microballoons by phase separation
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/08Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14Hydrolases (3)
    • C12N9/24Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405Glucanases
    • C12N9/2434Glucanases acting on beta-1,4-glucosidic bonds
    • C12N9/2437Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14Hydrolases (3)
    • C12N9/24Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405Glucanases
    • C12N9/2434Glucanases acting on beta-1,4-glucosidic bonds
    • C12N9/2445Beta-glucosidase (3.2.1.21)
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y302/00Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01004Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y302/00Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01021Beta-glucosidase (3.2.1.21)
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y302/00Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01091Cellulose 1,4-beta-cellobiosidase (3.2.1.91)

Abstract

The invention discloses a kind of carrier material fixed for cellulase and preparation method thereof, comprise the following steps:Obtained polyacrylonitrile complex microsphere is added in DMAC, after soaking 4 6h, after addition lithium chloride is well mixed, add chitin and catalyst, after well mixed, nitrogen is passed through, is heated after 2 4h are stirred under normal temperature, temperature is controlled at 100 120 DEG C, the 24h of stirring reaction 18, after being down to room temperature, after washes of absolute alcohol, cleaned again with deionized water, vacuumizing and drying.The carrier material mechanical strength of the application is excellent, and stability is preferable, stronger with the adhesion of enzyme, and enzyme molecule is not easy to come off from carrier in use, and service life is longer.Present invention also provides the application method of the carrier material fixed for cellulase.

Description

A kind of carrier material fixed for cellulase and preparation method thereof
Technical field
The present invention relates to enzymic preparation field, more particularly to a kind of carrier material fixed for cellulase and its preparation Method.
Background technology
Cellulase is a kind of multi-component compound enzyme system, mainly including circumscribed 1,4 beta-glucanase, inscribe beta glucan Enzyme and beta-glucosidase, by each component enzyme synergy by cellulose degradation into glucose.Each component of cellulase is big Part is glycoprotein, and the glycosyl in structure has certain protective effect from protease hydrolytic to cellulase.Separate sources Cellulase Molecules structure and folding mode are different, and their general character is generally by three parts structure composition:Spherical Catalytic activity area with catalysis, the land with cellulose-binding function and connect their one end height glycosyl The peptide acid of change.Current research thinks that cellulosic material will be digested effectively, needs cellulase being adsorbed in solid first On substrate, substrate cleavage is then being hydrolyzed into cell-oligosaccharide until glucose into small fragment.
Enzyme immobilizatio is that enzyme is incorporated on solid material or is limited in certain space, remains to maintain vigour and special Property, and a kind of technology that can be recycled.The preparation method of immobilised enzymes mainly has four kinds:Investment, cross-linking method, absorption Method and covalent coupling method.Investment, which refers to be embedded in enzyme molecule in the three-D space structure of porous polymer materials, consolidates enzyme Fixedization, this method preparation technology is simple, and influence of the formation condition gently and to enzymatic activity is relatively low, but the pore size of carrier Diffusion that can be to each material in enzymatic reaction has an impact.Cross-linking method is to react to be formed altogether with amino acid residue using crosslinking agent Valence link, enzyme molecule is set to be incorporated into network structure, so as to reach the effect of enzyme immobilization.Cross-linking method can make immobilised enzymes Stability improves, and service life extends, but covalent cross-linking reduces the reaction site of enzyme, so as to destroy the activity of enzyme.Absorption Method refers to that resolvase is incorporated on carrier by electrostatic interaction or hydrogen bond etc. to prepare the method for immobilised enzymes, and this method has can The advantages such as inverse property, simplification and enzyme activity height, but because the interaction of enzyme and carrier is weaker, be easy to come off.Covalent bond Method is that enzyme is fixed on suitable carrier by the method for Covalent bonding together, and enzyme produces chemical action with carrier, and adhesion is strong, Enzyme slip is low, has preferable stability and recycles performance, but reacts fierce, easily loses enzyme activity.The above four Kind method respectively has advantage and disadvantage, a variety of methods can be used to be used in mixed way to improve the use quality of immobilised enzymes.
Chinese patent CN201310674347.x discloses a kind of preparation method of cellulase immobilization carrier, including such as Lower step:(1) charing of cellulose, cellulose is carbonized under the conditions of 400 DEG C -500 DEG C of inert gas shielding;(2) carrier Modification, in the cellulose after charing, add ionic liquid and inorganic agent, stir, isothermal holding obtains cellulose carrier Solution;(3) carrier drying is molded, and solvent is added in cellulose carrier solution, and the washing that stirs removes impurity, Ran Hougan Cellulase immobilization carrier is made in dry, vacuum outgas.The invention has that carrier surface area is big, adsorption site is more, can be steady with enzyme Surely the advantages that combining.
The content of the invention
It is an object of the invention to provide a kind of carrier material fixed for cellulase and preparation method thereof, the carrier material Expect that mechanical strength is excellent, stability is preferable, stronger with the adhesion of enzyme, and enzyme molecule is not easy to take off from carrier in use Fall, service life is longer.
It is a further object of the present invention to provide the application method that this is used for the carrier material that cellulase is fixed.
To achieve the above object, the present invention uses following technical scheme:
A kind of preparation method for the carrier material fixed for cellulase, comprises the following steps:
(1) silester is dissolved in ethanol solution, adds nanometer Fe3O4Particle ultrasonic disperse 20-45min, Zhi Hou The mixture that glacial acetic acid and absolute ethyl alcohol are added dropwise in process is stirred vigorously, continues to stir 4-5h after being added dropwise to complete, stands 60- afterwards 72h, drying, Nano-meter SiO_2 is made after grinding2/Fe3O4Composite;
(2) polyacrylonitrile is dissolved in DMSO, adds Nano-meter SiO_22/Fe3O4Composite, ultrasonic disperse 1-2h, add Silane coupler is uniformly mixed, and is sprayed afterwards, and polyacrylonitrile complex microsphere is made;
(3) polyacrylonitrile complex microsphere made from step (2) is added in DMAC, after soaking 4-6h, adds lithium chloride and mix After closing uniformly, chitin and catalyst are added, after being well mixed, nitrogen is passed through, is heated after stirring 2-4h under normal temperature, control temperature At 100-120 DEG C, stirring reaction 18-24h, after being down to room temperature, clean, take out true after washes of absolute alcohol, then with deionized water Sky drying.
Preferably, nanometer Fe in the step (1)3O4The use quality of particle and the volume ratio of silester are 30-45g/ L。
Preferably, polyacrylonitrile and Nano-meter SiO_2 in the step (2)2/Fe3O4The mass ratio of composite is 4:1.
Preferably, the particle diameter of polyacrylonitrile complex microsphere prepared by the step (2) is 5 μm -7 μm.
Preferably, the usage amount mass ratio 5 of polyacrylonitrile complex microsphere and chitin in the step (3):28.
Preferably, catalyst is metallic sodium in the step (3).
A kind of carrier material fixed for cellulase, is prepared by above method.
A kind of immobilized cellulase, using it is described for cellulase fix carrier material as carrier, using glutaraldehyde as Crosslinking agent is prepared.
The invention has the advantages that
The application is with nanometer Fe3O4Nano-meter SiO_2 is made for core2/Fe3O4Composite, composite tool are magnetic same When there is more aperture, higher specific surface area, be a kind of mesoporous material of the magnetic carried.By Nano-meter SiO_22/Fe3O4It is multiple Condensation material is the polyacrylonitrile complex microsphere that filler is prepared into, and the specific surface area of complex microsphere can be effectively improved, so as to carry The high contact surface of polyacrylonitrile complex microsphere and cellulase, is relatively beneficial to improve load capacity of the cellulase on carrier, The load fastness of cellulase is improved, it is difficult for drop-off in use, improve organic efficiency.
Embodiment
In order to be better understood from the present invention, below by embodiment, the present invention is further described, and embodiment is served only for solving The present invention is released, any restriction will not be formed to the present invention.
Embodiment 1
A kind of preparation method for the carrier material fixed for cellulase, comprises the following steps:
(1) silester is dissolved in ethanol solution, adds nanometer Fe3O4Particle ultrasonic disperse 20min, nanometer Fe3O4 The use quality of particle and the volume ratio of silester are 30g/L, and glacial acetic acid and anhydrous is added dropwise in process is stirred vigorously afterwards The mixture of ethanol, continue to stir 4h after being added dropwise to complete, stand 60h afterwards, dry, Nano-meter SiO_2 is made after grinding2/Fe3O4It is multiple Condensation material;
(2) polyacrylonitrile is dissolved in DMSO, adds Nano-meter SiO_22/Fe3O4Composite, polyacrylonitrile and nanometer SiO2/Fe3O4The mass ratio of composite is 4:1, ultrasonic disperse 1h, add silane coupler and be uniformly mixed, carry out afterwards Spraying, polyacrylonitrile complex microsphere is made, the particle diameter of polyacrylonitrile complex microsphere is 5 μm -7 μm;
(3) polyacrylonitrile complex microsphere made from step (2) is added in DMAC, after soaking 4h, adds lithium chloride mixing After uniformly, the usage amount mass ratio 5 of chitin and catalyst metals sodium, polyacrylonitrile complex microsphere and chitin is added:28, mix After closing uniformly, nitrogen being passed through, is heated after stirring 2h under normal temperature, control temperature is at 100 DEG C, stirring reaction 18h, after being down to room temperature, Cleaned after washes of absolute alcohol, then with deionized water, vacuumizing and drying.
Using the carrier material fixed for cellulase as carrier, immobilization fibre is prepared with glutaraldehyde as cross linker Plain enzyme.
After tested, the immobilized cellulase prepared after 20 secondary responses its enzyme activity remain to keep original enzyme activity The 80% of power.
Embodiment 2
A kind of preparation method for the carrier material fixed for cellulase, comprises the following steps:
(1) silester is dissolved in ethanol solution, adds nanometer Fe3O4Particle ultrasonic disperse 45min, nanometer Fe3O4 The use quality of particle and the volume ratio of silester are 45g/L, and glacial acetic acid and anhydrous is added dropwise in process is stirred vigorously afterwards The mixture of ethanol, continue to stir 5h after being added dropwise to complete, stand 72h afterwards, dry, Nano-meter SiO_2 is made after grinding2/Fe3O4It is multiple Condensation material;
(2) polyacrylonitrile is dissolved in DMSO, adds Nano-meter SiO_22/Fe3O4Composite, polyacrylonitrile and nanometer SiO2/Fe3O4The mass ratio of composite is 4:1, ultrasonic disperse 2h, add silane coupler and be uniformly mixed, carry out afterwards Spraying, polyacrylonitrile complex microsphere is made, the particle diameter of polyacrylonitrile complex microsphere is 5 μm -7 μm;
(3) polyacrylonitrile complex microsphere made from step (2) is added in DMAC, after soaking 6h, adds lithium chloride mixing After uniformly, the usage amount mass ratio 5 of chitin and catalyst metals sodium, polyacrylonitrile complex microsphere and chitin is added:28, mix After closing uniformly, nitrogen being passed through, is heated after stirring 4h under normal temperature, control temperature is at 120 DEG C, stirring reaction 24h, after being down to room temperature, Cleaned after washes of absolute alcohol, then with deionized water, vacuumizing and drying.
Using the carrier material fixed for cellulase as carrier, immobilization fibre is prepared with glutaraldehyde as cross linker Plain enzyme.
After tested, the immobilized cellulase prepared after 20 secondary responses its enzyme activity remain to keep original enzyme activity The 82% of power.
Embodiment 3
A kind of preparation method for the carrier material fixed for cellulase, comprises the following steps:
(1) silester is dissolved in ethanol solution, adds nanometer Fe3O4Particle ultrasonic disperse 20min, nanometer Fe3O4 The use quality of particle and the volume ratio of silester are 45g/L, and glacial acetic acid and anhydrous is added dropwise in process is stirred vigorously afterwards The mixture of ethanol, continue to stir 4h after being added dropwise to complete, stand 72h afterwards, dry, Nano-meter SiO_2 is made after grinding2/Fe3O4It is multiple Condensation material;
(2) polyacrylonitrile is dissolved in DMSO, adds Nano-meter SiO_22/Fe3O4Composite, polyacrylonitrile and nanometer SiO2/Fe3O4The mass ratio of composite is 4:1, ultrasonic disperse 1h, add silane coupler and be uniformly mixed, carry out afterwards Spraying, polyacrylonitrile complex microsphere is made, the particle diameter of polyacrylonitrile complex microsphere is 5 μm -7 μm;
(3) polyacrylonitrile complex microsphere made from step (2) is added in DMAC, after soaking 6h, adds lithium chloride mixing After uniformly, the usage amount mass ratio 5 of chitin and catalyst metals sodium, polyacrylonitrile complex microsphere and chitin is added:28, mix After closing uniformly, nitrogen being passed through, is heated after stirring 2h under normal temperature, control temperature is at 120 DEG C, stirring reaction 18h, after being down to room temperature, Cleaned after washes of absolute alcohol, then with deionized water, vacuumizing and drying.
Using the carrier material fixed for cellulase as carrier, immobilization fibre is prepared with glutaraldehyde as cross linker Plain enzyme.
After tested, the immobilized cellulase prepared after 20 secondary responses its enzyme activity remain to keep original enzyme activity The 89% of power.
Embodiment 4
A kind of preparation method for the carrier material fixed for cellulase, comprises the following steps:
(1) silester is dissolved in ethanol solution, adds nanometer Fe3O4Particle ultrasonic disperse 45min, nanometer Fe3O4 The use quality of particle and the volume ratio of silester are 30g/L, and glacial acetic acid and anhydrous is added dropwise in process is stirred vigorously afterwards The mixture of ethanol, continue to stir 5h after being added dropwise to complete, stand 60h afterwards, dry, Nano-meter SiO_2 is made after grinding2/Fe3O4It is multiple Condensation material;
(2) polyacrylonitrile is dissolved in DMSO, adds Nano-meter SiO_22/Fe3O4Composite, polyacrylonitrile and nanometer SiO2/Fe3O4The mass ratio of composite is 4:1, ultrasonic disperse 2h, add silane coupler and be uniformly mixed, carry out afterwards Spraying, polyacrylonitrile complex microsphere is made, the particle diameter of polyacrylonitrile complex microsphere is 5 μm -7 μm;
(3) polyacrylonitrile complex microsphere made from step (2) is added in DMAC, after soaking 4h, adds lithium chloride mixing After uniformly, the usage amount mass ratio 5 of chitin and catalyst metals sodium, polyacrylonitrile complex microsphere and chitin is added:28, mix After closing uniformly, nitrogen being passed through, is heated after stirring 4h under normal temperature, control temperature is at 100 DEG C, stirring reaction 24h, after being down to room temperature, Cleaned after washes of absolute alcohol, then with deionized water, vacuumizing and drying.
Using the carrier material fixed for cellulase as carrier, immobilization fibre is prepared with glutaraldehyde as cross linker Plain enzyme.
After tested, the immobilized cellulase prepared after 20 secondary responses its enzyme activity remain to keep original enzyme activity The 88% of power.
Embodiment 5
A kind of preparation method for the carrier material fixed for cellulase, comprises the following steps:
(1) silester is dissolved in ethanol solution, adds nanometer Fe3O4Particle ultrasonic disperse 30min, nanometer Fe3O4 The use quality of particle and the volume ratio of silester are 40g/L, and glacial acetic acid and anhydrous is added dropwise in process is stirred vigorously afterwards The mixture of ethanol, continue to stir 5h after being added dropwise to complete, stand 66h afterwards, dry, Nano-meter SiO_2 is made after grinding2/Fe3O4It is multiple Condensation material;
(2) polyacrylonitrile is dissolved in DMSO, adds Nano-meter SiO_22/Fe3O4Composite, polyacrylonitrile and nanometer SiO2/Fe3O4The mass ratio of composite is 4:1, ultrasonic disperse 2h, add silane coupler and be uniformly mixed, carry out afterwards Spraying, polyacrylonitrile complex microsphere is made, the particle diameter of polyacrylonitrile complex microsphere is 5 μm -7 μm;
(3) polyacrylonitrile complex microsphere made from step (2) is added in DMAC, after soaking 5h, adds lithium chloride mixing After uniformly, the usage amount mass ratio 5 of chitin and catalyst metals sodium, polyacrylonitrile complex microsphere and chitin is added:28, mix After closing uniformly, nitrogen being passed through, is heated after stirring 3h under normal temperature, control temperature is at 110 DEG C, stirring reaction 20h, after being down to room temperature, Cleaned after washes of absolute alcohol, then with deionized water, vacuumizing and drying.
Using the carrier material fixed for cellulase as carrier, immobilization fibre is prepared with glutaraldehyde as cross linker Plain enzyme.
After tested, the immobilized cellulase prepared after 20 secondary responses its enzyme activity remain to keep original enzyme activity The 85% of power.

Claims (8)

1. a kind of preparation method for the carrier material fixed for cellulase, it is characterised in that comprise the following steps:
(1) silester is dissolved in ethanol solution, adds nanometer Fe3O4Particle ultrasonic disperse 20-45min, afterwards violent The mixture of glacial acetic acid and absolute ethyl alcohol is added dropwise in whipping process, continues to stir 4-5h after being added dropwise to complete, stands 60-72h afterwards, Drying, Nano-meter SiO_2 is made after grinding2/Fe3O4Composite;
(2) polyacrylonitrile is dissolved in DMSO, adds Nano-meter SiO_22/Fe3O4Composite, ultrasonic disperse 1-2h, add silane Coupling agent is uniformly mixed, and is sprayed afterwards, and polyacrylonitrile complex microsphere is made;
(3) polyacrylonitrile complex microsphere made from step (2) is added in DMAC, after soaking 4-6h, it is equal adds lithium chloride mixing After even, chitin and catalyst are added, after being well mixed, nitrogen is passed through, is heated after stirring 2-4h under normal temperature, control temperature exists 100-120 DEG C, stirring reaction 18-24h, after being down to room temperature, clean, vacuumize after washes of absolute alcohol, then with deionized water Drying.
2. the preparation method of the carrier material according to claim 1 fixed for cellulase, it is characterised in that:It is described Nanometer Fe in step (1)3O4The use quality of particle and the volume ratio of silester are 30-45g/L.
3. the preparation method of the carrier material according to claim 1 fixed for cellulase, it is characterised in that:It is described Polyacrylonitrile and Nano-meter SiO_2 in step (2)2/Fe3O4The mass ratio of composite is 4:1.
4. the preparation method of the carrier material according to claim 1 fixed for cellulase, it is characterised in that:It is described The particle diameter of polyacrylonitrile complex microsphere prepared by step (2) is 5 μm -7 μm.
5. the preparation method of the carrier material according to claim 1 fixed for cellulase, it is characterised in that:It is described The usage amount mass ratio 5 of polyacrylonitrile complex microsphere and chitin in step (3):28.
6. the preparation method of the carrier material according to claim 1 fixed for cellulase, it is characterised in that:It is described Catalyst is metallic sodium in step (3).
A kind of 7. carrier material fixed for cellulase, it is characterised in that:As the method described in claim any one of 1-6 It is prepared.
A kind of 8. immobilized cellulase, it is characterised in that:With the carrier material for being used for cellulase and fixing described in claim 7 Expect for carrier, to be prepared with glutaraldehyde as cross linker.
CN201710639092.1A 2017-07-31 2017-07-31 A kind of carrier material fixed for cellulase and preparation method thereof Pending CN107418949A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710639092.1A CN107418949A (en) 2017-07-31 2017-07-31 A kind of carrier material fixed for cellulase and preparation method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710639092.1A CN107418949A (en) 2017-07-31 2017-07-31 A kind of carrier material fixed for cellulase and preparation method thereof

Publications (1)

Publication Number Publication Date
CN107418949A true CN107418949A (en) 2017-12-01

Family

ID=60431445

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710639092.1A Pending CN107418949A (en) 2017-07-31 2017-07-31 A kind of carrier material fixed for cellulase and preparation method thereof

Country Status (1)

Country Link
CN (1) CN107418949A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109315795A (en) * 2018-11-27 2019-02-12 湖北文理学院 A kind of method of enzyme process joint infra-red drying removal walnut kernel pellicle

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1296032A (en) * 2000-11-21 2001-05-23 上海博纳科技发展有限公司 Polyacrylonitrile magnetic pearl and preparation process and application thereof
CN1978635A (en) * 2005-12-06 2007-06-13 上海师范大学 Magnetic nano artificial incision enzyme, and its preparing method and use
CN101012312A (en) * 2007-02-08 2007-08-08 上海交通大学 Method of preparing multifunctional macromolecule-inorganic composite microsphere
US20090277843A1 (en) * 2008-05-08 2009-11-12 Kabushiki Kaisha Toshiba Polymer composite, water-treatment method using the same and manufacturing method of the same
CN104415364A (en) * 2013-08-27 2015-03-18 成都市绿科华通科技有限公司 Polyacrylonitrile-containing medical high-molecular contrast agent microballoon
CN106268810A (en) * 2016-07-29 2017-01-04 华南理工大学 A kind of surface recombination modifies particle aggregate carrier material and preparation method and application

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1296032A (en) * 2000-11-21 2001-05-23 上海博纳科技发展有限公司 Polyacrylonitrile magnetic pearl and preparation process and application thereof
CN1978635A (en) * 2005-12-06 2007-06-13 上海师范大学 Magnetic nano artificial incision enzyme, and its preparing method and use
CN101012312A (en) * 2007-02-08 2007-08-08 上海交通大学 Method of preparing multifunctional macromolecule-inorganic composite microsphere
US20090277843A1 (en) * 2008-05-08 2009-11-12 Kabushiki Kaisha Toshiba Polymer composite, water-treatment method using the same and manufacturing method of the same
CN104415364A (en) * 2013-08-27 2015-03-18 成都市绿科华通科技有限公司 Polyacrylonitrile-containing medical high-molecular contrast agent microballoon
CN106268810A (en) * 2016-07-29 2017-01-04 华南理工大学 A kind of surface recombination modifies particle aggregate carrier material and preparation method and application

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
张猛等: "氨基硅烷化磁性纳米微球固定化纤维素酶研究", 《太阳能学报》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109315795A (en) * 2018-11-27 2019-02-12 湖北文理学院 A kind of method of enzyme process joint infra-red drying removal walnut kernel pellicle

Similar Documents

Publication Publication Date Title
Chang et al. Cellulase immobilized mesoporous silica nanocatalysts for efficient cellulose-to-glucose conversion
CN104448397B (en) A kind of cellulose-silicon dioxide composite aerogel in-situ preparation method
CN103723824B (en) Carbon-iron microbattery slow-release carbon source filler and preparation method thereof
CN105642233B (en) A kind of method that continuity method prepares CMC/GO composite hydrogel microballoons
Yang Uniform cross-linked cellulase aggregates prepared in millifluidic reactors
CN103710333A (en) Carrier for immobilization as well as preparation method thereof and immobilized beta-glucosaccharase
CN107893062B (en) Method for immobilizing cellulase and hydrolyzing cellulose
CN102952792B (en) Preparation method of beta-glucosidase immobilized by sodium alginate loaded with attapulgite
CN111672432A (en) Preparation method of graphene oxide/chitosan composite aerogel for enzyme immobilization
CN106752833A (en) A kind of preparation method of graphene oxide/silica composite coating
CN107418949A (en) A kind of carrier material fixed for cellulase and preparation method thereof
CN109201029B (en) Preparation method of efficient porous composite photocatalytic material
CN101671663B (en) Preparation method of corncob fixed lipase and product thereof
CN106179295B (en) A kind of photocatalytic activity Carbon fibe and preparation method thereof
CN105038500A (en) Urushiol-modified aerosil super thermal-insulation heat-preserving paint and preparation method thereof
CN103265720B (en) Novel method for preparing porous crosslinked chitosan microsphere
CN101096665A (en) Application of halloysite in enzyme immobilization carrier
CN101818139A (en) Preparation method of immobilization peroxidase
CN103013975B (en) Method for preparing CaSiO<3>@SiO<2> particles containing immobilized enzymes
CN104531665A (en) Functionalized active attapulgite enzyme immobilization catalyst preparation method
CN107338238B (en) Immobilized laccase and preparation method thereof
CN111495330B (en) High-specific-surface-area multi-dimensional biochar adsorption matrix and preparation method thereof
CN104888531A (en) Preparation process of nano composite filter material of gas turbine device
CN109999857B (en) Near-infrared response hollow cerium fluoride up-conversion photocatalytic material and preparation method and application thereof
CN116983953B (en) Preparation method of aerogel microspheres

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
WD01 Invention patent application deemed withdrawn after publication
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20171201