CN101892216A - Method for preparing carrier material for use in amino-acylase immobilization by polyamidoamine dendrimer - Google Patents
Method for preparing carrier material for use in amino-acylase immobilization by polyamidoamine dendrimer Download PDFInfo
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- CN101892216A CN101892216A CN2009100844979A CN200910084497A CN101892216A CN 101892216 A CN101892216 A CN 101892216A CN 2009100844979 A CN2009100844979 A CN 2009100844979A CN 200910084497 A CN200910084497 A CN 200910084497A CN 101892216 A CN101892216 A CN 101892216A
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Abstract
The invention relates to a method for preparing a novel immobilized enzyme carrier material, in particular to a method for preparing an immobilized enzyme carrier material which uses dendrimer modified-surface silica gel as a substrate and amino-acylase as a cross-linking ligand. A divergent synthesis method is used to bond the dendrimer onto the surface of the silica gel to prepare the dendrimer modified-surface silica gel. The cross linking between the dendrimer modified-surface silica gel and the amino-acylase is realized by a glutaraldehyde crosslinking method, and the carrier material is used for the separation of N-acetyl-DL-methionine.
Description
Technical field
The present invention relates to a kind of preparation method of novel Carrier Materials of Immobilized Enzyme, is matrix with dendrimer finishing silica gel particularly, with the preparation method of L-Aminoacylase as the Carrier Materials of Immobilized Enzyme of crosslinking ligand.
Background technology
Dendritic macromole (Dendrimer) is the novel functional polymer compound of a class, and its feature is to carry out the design of bulk of molecule, shape and functional group at molecular level.Because its unique texture has a wide range of applications in fields such as medical science, Materials science, life science and analytical chemistry.Dendritic macromole is guided to the silica gel surface, can increase surface-area, capacity and the selectivity of silica gel greatly.Immobilized enzyme has good stability, can repeat and advantage such as use continuously, and can improve degree of purity of production and process efficiency, overcomes free biological catalyst to environment sensitive, character instability, easy shortcoming such as inactivation or death.Carrier has bigger influence to immobilized enzyme, make the ideal immobilized enzyme, should select the rational and effective process for fixation for use, and good carrier is arranged simultaneously again.Therefore the fixed enzyme vector by the synthetic excellent property of modern technologies is the main research contents of immobilized enzyme research field.
Summary of the invention
The objective of the invention is to study a kind of preparation method of novel Carrier Materials of Immobilized Enzyme; dendrimer is applied in the preparation of immobilized aminoacylase; can improve stability, repeatability and the life-span of immobilized enzyme greatly; simplify aftertreatment; improve yield and selectivity; thereby improve the work-ing life of immobilized enzyme, reduce the influence of reaction conditions, improve catalytic efficiency immobilized enzyme.End amino at dendritic macromole is submitted the hydrazino acylase, makes it reach the purpose of separating the N-acetyl-DL-amino acid by reacting with N-acetyl-L-amino acid.
Main points of the present invention are to utilize the 26S Proteasome Structure and Function characteristics of polyamide-amide dendrimer, earlier it are bonded to the silica gel surface, make polyamide-amide finishing silica gel, and continue to submit hydrazino acylase part at its end amino.Its concrete preparation process comprises:
(1) dendrimer finishing silica gel is synthetic: do silylating reagent with the 3-aminopropyl triethoxysilane, synthesis method is dispersed in employing, with methyl acrylate and quadrol is reactant, by repeated Markovnikov addition reaction, amidate action, obtained the silica gel of polyamide-amide finishing.
(2) L-Aminoacylase is crosslinked: utilize the glutaraldehyde cross-linking method that L-Aminoacylase is cross-linked on the end amino of the silica gel that dendritic macromole polyamide-amine modifies, synthetic is spacerarm with the polyamide-amide, and L-Aminoacylase is the novel immobilized enzyme of part.
Description of drawings
1, the building-up process of Fig. 1 polyamide-amide dendrimer finishing silica gel
2, the cross-linking process of Fig. 2 L-Aminoacylase
Embodiment
Synthesizing of dendrimer finishing silica gel:
Silica gel is placed the 50ml round-bottomed flask, add 1molL
-1Aqueous hydrochloric acid, the following 100 ℃ of back flow reaction of induction stirring 6 hours; Leach solid, it is neutral washing repeatedly to water lotion with deionized water, and vacuum-drying is to constant weight under the room temperature.Mix with toluene then and stirred 10 minutes, afterwards the toluene solution of silica gel, 10% 3-aminopropyl triethoxysilane added and place the 50ml round-bottomed flask, stirring at room 24 hours, reflux at 120 ℃ again and stirred 2.5 hours, be cooled to suction filtration after the room temperature, leach thing for several times, at 47 ℃ of vacuum-drying 24h with methanol wash.
The silica gel and the methanol mixed in last step were stirred 10 minutes, slowly add 20% methyl acrylate methanol solution then,, use washed with methanol 3-4 time afterwards 25 ℃ of isothermal reactions 12.5 hours.47 ℃ of vacuum-dryings 24 hours, obtain the polyamide-amide finishing silica gel in 0.5 generation.The silica gel and the methanol mixed of bonding 0.5 generation polyamide-amide were stirred 10 minutes, slowly add 20% quadrol methanol solution then,, use washed with methanol 3-4 time then 40 ℃ of isothermal reactions 19.5 hours.47 ℃ of vacuum-dryings 24 hours, obtain the polyamide-amide finishing silica gel in 1.0 generations.Repeat above Markovnikov addition reaction and amidate action, the polyamide-amide finishing silica gel in synthetic respectively 1.5 generations, 2.0 generations, 2.5 generations, 3.0 generations.
Embodiment 2:
L-Aminoacylase crosslinked:
With through the silica gel of 3.0 generations polyamide-amide finishing at normal temperatures with 1% glutaraldehyde cross-linking 4 hours, filter the back and wash repeatedly with deionized water and remove excessive glutaraldehyde, 30 ℃ of vacuum-dryings 12 hours; Afterwards reacted silica gel is joined 0.2molL
-1L-Aminoacylase solution in (solution leaves standstill after 30 minutes and uses with the preparation of the phosphoric acid buffer of pH=7.0), reacted at normal temperatures 2 hours, filter the back with deionized water rinsing 5-6 time, seal standby.
Claims (3)
1. the preparation method of a novel Carrier Materials of Immobilized Enzyme; it is characterized in that: do silylating reagent with the 3-aminopropyl triethoxysilane; synthesis method is dispersed in employing; with methyl acrylate and quadrol is reactant; by repeated Markovnikov addition reaction, amidate action; obtained the silica gel of polyamide-amide finishing; and utilize the glutaraldehyde cross-linking method L-Aminoacylase to be cross-linked on the end amino of the silica gel that dendritic macromole polyamide-amine modifies; obtaining with the polyamide-amide is spacerarm, and L-Aminoacylase is the novel immobilized enzyme of part.
2. the preparation of the described novel Carrier Materials of Immobilized Enzyme of preparation claim 1 is characterized in that: the preparation process that dendrimer is applied to the polyamide-amide finishing silica gel of Carrier Materials of Immobilized Enzyme.
3. according to the preparation of the described novel Carrier Materials of Immobilized Enzyme of claim 1, it is characterized in that: further bonding L-Aminoacylase on the silica gel of the polyamide-amide finishing of gained.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100844979A CN101892216A (en) | 2009-05-20 | 2009-05-20 | Method for preparing carrier material for use in amino-acylase immobilization by polyamidoamine dendrimer |
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| Application Number | Priority Date | Filing Date | Title |
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| CN2009100844979A CN101892216A (en) | 2009-05-20 | 2009-05-20 | Method for preparing carrier material for use in amino-acylase immobilization by polyamidoamine dendrimer |
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| CN101892216A true CN101892216A (en) | 2010-11-24 |
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| CN2009100844979A Pending CN101892216A (en) | 2009-05-20 | 2009-05-20 | Method for preparing carrier material for use in amino-acylase immobilization by polyamidoamine dendrimer |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102161758A (en) * | 2011-01-26 | 2011-08-24 | 大连理工大学 | Preparation method of novel silica gel-based hyperbranched PAMAM (polyamidoamine) chelating resin |
| CN104558628A (en) * | 2013-10-17 | 2015-04-29 | 中国石油化工股份有限公司 | Magnetic nanoscale resin compound as well as preparation method and application thereof |
| CN110183667A (en) * | 2019-06-24 | 2019-08-30 | 南京大学 | A kind of preparation method and applications of the without phosphorus broom shaped polymer of silica gel load |
| CN110394148A (en) * | 2019-07-05 | 2019-11-01 | 重庆科技学院 | It is a kind of with the composite material and preparation method thereof of methyl acrylate modified silica-gel and product and application |
-
2009
- 2009-05-20 CN CN2009100844979A patent/CN101892216A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102161758A (en) * | 2011-01-26 | 2011-08-24 | 大连理工大学 | Preparation method of novel silica gel-based hyperbranched PAMAM (polyamidoamine) chelating resin |
| CN104558628A (en) * | 2013-10-17 | 2015-04-29 | 中国石油化工股份有限公司 | Magnetic nanoscale resin compound as well as preparation method and application thereof |
| CN110183667A (en) * | 2019-06-24 | 2019-08-30 | 南京大学 | A kind of preparation method and applications of the without phosphorus broom shaped polymer of silica gel load |
| CN110183667B (en) * | 2019-06-24 | 2021-07-16 | 南京大学 | Preparation method and application of silica gel loaded phosphorus-free broom-like polymer |
| CN110394148A (en) * | 2019-07-05 | 2019-11-01 | 重庆科技学院 | It is a kind of with the composite material and preparation method thereof of methyl acrylate modified silica-gel and product and application |
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Application publication date: 20101124 |