CN1376792A - Process for preparing magnetic microsphere immobilized cellulase - Google Patents
Process for preparing magnetic microsphere immobilized cellulase Download PDFInfo
- Publication number
- CN1376792A CN1376792A CN 01110024 CN01110024A CN1376792A CN 1376792 A CN1376792 A CN 1376792A CN 01110024 CN01110024 CN 01110024 CN 01110024 A CN01110024 A CN 01110024A CN 1376792 A CN1376792 A CN 1376792A
- Authority
- CN
- China
- Prior art keywords
- cellulase
- magnetic
- plain
- immobilized cellulase
- magnetic 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.)
- Granted
Links
Images
Landscapes
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Abstract
A process for preparing the cellulase immobilized an magentic microphseres features that the industrial cellulase is physically adsorbed by magentic agarose micropheres. Its resultant cellulase features magentic response, high content of carried protein, high specific activity, high activity recovering rate and low cost.
Description
The present invention relates to a kind of preparation method of magnetic microsphere immobilized cellulase, it comprises physical adsorption process.
Magnetic macromolecular microsphere can be in conjunction with various functional moleculars, as enzyme, antibody, cell, DNA etc., thereby are used widely in fields such as cytobiology, molecular biology, medical science.As US5053332, the preparation method of the sepharose 4B that mixes biomaterial is promptly disclosed.This method is mainly used in the embedding of cell.
The preparation method who the purpose of this invention is to provide a kind of magnetic microsphere immobilized cellulase, present method is mixed through physical adsorption with cellulase with the magnetic agarose microbeads and is prepared, the magnetic microsphere immobilized cellulase that present method is made, its activity recovery can reach more than 70%, compare with natural enzyme, heat, ph stability are all increased.
The objective of the invention is to finish by following measure:
With the plain zymin adding distil water of industrial fiber uniform mixing in beaker, the ratio of plain enzyme of industrial fiber and distilled water is 1g:20～100ml, filter through dusting cover, filtrate adds solid ammonium sulfate to 20% degree of saturation, with 2000～6000 rev/mins, centrifugal 5～30 minutes, abandon precipitation, in supernatant liquor, add the magnetic agarose microbeads, the consumption mass ratio of plain zymin of industrial fiber and microballoon is 1: 1～10, add solid ammonium sulfate to 60% saturation ratio behind the mixing again, 4～10 ℃ of absorption 1～24 hour of vibrating down, beaker is transferred permanent magnet, magneticstrength＞0.04T, precipitation is abandoned supernatant liquor after the sedimentation, be washed till no zymoprotein with distilled water again till, gained precipitates and is magnetic microsphere immobilized cellulase.
Advantage of the present invention is:
Present method is a carrier with the magnetic agarose microbeads, by physical adsorption the plain zymin of industrial fiber is fixed on the microballoon, is prepared into the magnetic microsphere immobilized cellulase with magnetic responsiveness.This product has high protein carrying capacity, alive, the high reactivity rate of recovery of height ratio.Present method just can make things convenient for by imposing the external magnetic field, quickly and easily enzyme be reclaimed, uses repeatedly, thereby has improved the utilization ratio of cellulase, has reduced cost.This preparation process need not to adopt high temperature, organic solvent or complicated production unit.This product application all can produce tangible economic benefit in the production of foodstuffs industry, fermentation industry, cellulosic saccharification, fuel alcohol.
Adsorption time is to the influence of magnetic immobilized cellulase:
Parallel each 5 parts of the plain zymin 10g of magnetic composite agarose microspheres 11.8g and industrial fiber that get, constant other conditions are reacted under different time, the results are shown in Table 1.Table 1 adsorption time is to the influence of magnetic immobilized cellulase
| Adsorption time/h | Total activity/u/g microballoon | Albumen carrying capacity/mg/g microballoon | Than work/U/mg albumen | Activity recovery 1% |
| ????1 | ????158.3 | ????95.8 | ????1.65 | ????63.5 |
| ????2 | ????183.3 | ????106.3 | ????1.72 | ????65.4 |
| ????8 | ????191.7 | ????100.0 | ????1.92 | ????73.1 |
| ????12 | ????175.0 | ????125.0 | ????1.40 | ????53.8 |
| ????24 | ????170.8 | ????116.0 | ????1.47 | ????57.7 |
The operational stability of magnetic immobilized cellulase:
Get the carboxymethylcellulose sodium solution 20ml of magnetic immobilized cellulase 0.2g and 10% (massfraction),, survey its vigor with the DNS method at PH5.0 conditioned response 30min.So reuse 10 times, its active maintenance situation sees Table 2.
Table 2 magnetic immobilized cellulase operational stability
| The enzyme access times | ???1 | ???2 | ???3 | ???4 | ???5 | ???6 | ???7 | ???8 | ???9 |
| Relative vigor % | ?100 | ??94 | ??88 | ??84 | ??84 | ??72 | ??70 | ??64 | ??60 |
This accompanying drawing is preparation technology's schema:
Embodiment:
Get the plain zymin adding distil water of 10g industrial fiber 500ml, uniform mixing in beaker, filter through dusting cover, filtrate adds solid ammonium sulfate to 20% degree of saturation, with 4000 rev/mins, centrifugal 20 minutes, abandon precipitation, in supernatant liquor, add 11.8g magnetic agarose microbeads, add solid ammonium sulfate to 60% saturation ratio behind the mixing again, vibrated 8 hours down at 6 ℃, beaker is transferred permanent magnet, and magneticstrength is 0.05T, abandons supernatant liquor after the precipitation sedimentation, till being washed till no zymoprotein with distilled water again, the gained precipitation is magnetic microsphere immobilized cellulase.The albumen carrying capacity of this magnetic enzyme immobilization cellulase is the 100mg/g microballoon, is that 1.92U/mg albumen, activity recovery are 73.1% than living.
Claims (1)
1. the preparation method of magnetic microsphere immobilized cellulase, it comprises physical adsorption process, it is characterized in that with the plain zymin adding distil water of industrial fiber uniform mixing in beaker, the ratio of plain enzyme of industrial fiber and distilled water is 1g:20～100ml, filter through dusting cover, filtrate adds solid ammonium sulfate to 20% degree of saturation, with 2000～6000 rev/mins, centrifugal 5～30 minutes, abandon precipitation, in supernatant liquor, add the magnetic agarose microbeads, the consumption mass ratio of plain zymin of industrial fiber and microballoon is 1: 1～10, add solid ammonium sulfate to 60% saturation ratio behind the mixing again, 4～10 ℃ of absorption 1～24 hour of vibrating down, beaker is transferred permanent magnet, magneticstrength＞0.04T, precipitation is abandoned supernatant liquor after the sedimentation, be washed till no zymoprotein with distilled water again till, gained precipitates and is magnetic microsphere immobilized cellulase.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01110024 CN1224703C (en) | 2001-03-27 | 2001-03-27 | Process for preparing magnetic microsphere immobilized cellulase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01110024 CN1224703C (en) | 2001-03-27 | 2001-03-27 | Process for preparing magnetic microsphere immobilized cellulase |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1376792A true CN1376792A (en) | 2002-10-30 |
| CN1224703C CN1224703C (en) | 2005-10-26 |
Family
ID=4658292
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 01110024 Expired - Fee Related CN1224703C (en) | 2001-03-27 | 2001-03-27 | Process for preparing magnetic microsphere immobilized cellulase |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1224703C (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101643727B (en) * | 2008-08-05 | 2011-04-20 | 中国农业大学 | Hollow polysaccharide microsphere immobilized enzyme and preparation method thereof |
| CN101643725B (en) * | 2008-08-05 | 2011-06-01 | 中国农业大学 | Magnetic hollow compound micro-structure immobilized enzyme and method for preparing same |
| CN1975429B (en) * | 2006-12-14 | 2011-11-02 | 复旦大学 | Nano-magnetic microsphere with amino having fixed protease at surface, preparing method and application thereof |
| CN102994489A (en) * | 2012-10-09 | 2013-03-27 | 山西农业大学 | Biotin-avidin system immobilized glucoamylase and preparation method thereof |
| CN109970879A (en) * | 2019-03-21 | 2019-07-05 | 浙江中医药大学 | A kind of bletilla polysaccharide extract and preparation method thereof |
-
2001
- 2001-03-27 CN CN 01110024 patent/CN1224703C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1975429B (en) * | 2006-12-14 | 2011-11-02 | 复旦大学 | Nano-magnetic microsphere with amino having fixed protease at surface, preparing method and application thereof |
| CN101643727B (en) * | 2008-08-05 | 2011-04-20 | 中国农业大学 | Hollow polysaccharide microsphere immobilized enzyme and preparation method thereof |
| CN101643725B (en) * | 2008-08-05 | 2011-06-01 | 中国农业大学 | Magnetic hollow compound micro-structure immobilized enzyme and method for preparing same |
| CN102994489A (en) * | 2012-10-09 | 2013-03-27 | 山西农业大学 | Biotin-avidin system immobilized glucoamylase and preparation method thereof |
| CN102994489B (en) * | 2012-10-09 | 2014-01-08 | 山西农业大学 | Biotin-avidin system immobilized glucoamylase and preparation method thereof |
| CN109970879A (en) * | 2019-03-21 | 2019-07-05 | 浙江中医药大学 | A kind of bletilla polysaccharide extract and preparation method thereof |
| CN109970879B (en) * | 2019-03-21 | 2022-03-08 | 浙江中医药大学 | Bletilla striata polysaccharide extract and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1224703C (en) | 2005-10-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Tan et al. | Production of xylobiose from the autohydrolysis explosion liquor of corncob using Thermotoga maritima xylanase B (XynB) immobilized on nickel-chelated Eupergit C | |
| CN101235368A (en) | Method for preparing immobilized enzyme biological catalyst | |
| CN103224926A (en) | Method of preparing immobilized lipase | |
| CN112553186B (en) | Copper-based metal organic framework material immobilized laccase and preparation method and application thereof | |
| CN109576256B (en) | Method for encapsulating double enzymes by magnetic DNA hydrogel | |
| CN101280298A (en) | Preparation of recyclable magnetic nanometer immobilized enzyme | |
| CN1224703C (en) | Process for preparing magnetic microsphere immobilized cellulase | |
| Rogalski et al. | Immobilization of cellulase and D-xylanase complexes from Aspergillus terreus F-413 on controlled porosity glasses | |
| CN113025678A (en) | Method for screening hypoglycemic peptide capable of inhibiting alpha-amylase activity | |
| CN101575594B (en) | Technique method of immobilized candida antarctica lipase B | |
| Ozyilmaz et al. | Preparation of regenerable magnetic nanoparticles for cellulase immobilization: Improvement of enzymatic activity and stability | |
| CN109207467A (en) | A kind of method for immobilizing cell of grate sulfur thiobacillus | |
| CN104293763B (en) | The method of lipase immobilization carrier and its fixed fat enzyme | |
| CN103509778B (en) | A kind of extracting and purifying method of halogenated acid dehalogenase | |
| CN106148319B (en) | Method for preparing immobilized enzyme based on reaction adsorption method | |
| CN104711248A (en) | Substrate toxicity eliminating method | |
| Zhu et al. | Immobilization of glycolate oxidase from Medicago falcata on magnetic nanoparticles for application in biosynthesis of glyoxylic acid | |
| CN105039299B (en) | A kind of immobilized HRP carrier and its preparation, application process | |
| CN114231521B (en) | Preparation method and application of modified attapulgite immobilized enzyme | |
| CN114657170B (en) | Preparation method of high-stability immobilized enzyme | |
| Sun et al. | Preparation of 3D porous cellulose‐chitosan hybrid gel macrospheres by alkaline urea system for enzyme immobilization | |
| CN101845465A (en) | Process for producing D(-)-tartaric acid by biotransformation | |
| CN107326021B (en) | Preparation method of magnetic cellulose microsphere immobilized lipase catalyst | |
| CN100383239C (en) | Process for extracting oxalate oxidase from bran of wheat | |
| CN112795598B (en) | Method for producing inositol based on silicon mineralized microcapsule immobilized multienzyme |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20051026 Termination date: 20100327 |