Unlock instant, AI-driven research and patent intelligence for your innovation.

A magnetically cross-linked lipase aggregate and its preparation method and application

A technique of lipase, aggregates, applied in the direction of immobilization on or in inorganic carriers, immobilization on/in organic carriers, etc.

Inactive Publication Date: 2016-04-20
INST OF PROCESS ENG CHINESE ACAD OF SCI
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, magnetic nanoparticles are rarely used to prepare magnetically cross-linked lipase aggregates, and there are few reports, especially the preparation of magnetic cross-linked lipase aggregates using functionalized magnetic nanoparticles with dendritic polyamine groups on the surface. No literature report

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] (1) Weigh 15 mg self-made functionalized magnetic nanoparticles with dendritic amine groups on the surface (preparation method: magnetic nanopolymers with surface amination (see: GuobinShan, HuizhouLiu, JianminXing, GuandongZhang, ZhiyaMa, XianqiaoLiu, JunguoLiu .SurfaceModificationofNano-magneticFe3O4ParticlesanditsApplications.China-EUForumonNanosizedTechnology, 2002, Beijing, China.P68-76.) as the core, with ethylenediamine and methyl acrylate as monomers, using a divergent synthesis method known to those skilled in the art to synthesize dendritic materials. ) placed in a conical flask filled with 100mM phosphate buffer solution of 15mlpH=8, ultrasonicated in an ice bath for 20min under ultrasonic conditions with a frequency of 40KHz and a power of 240w, and the dispersion was uniform;

[0042] (2) Weigh 250mg of lipase (based on protein content) and place it in 250ml of 100mM phosphate buffer, stir magnetically for 30min at room temperature to fully dissolve the lipa...

Embodiment 2

[0047] (1) Weigh 12mg of functionalized magnetic nanoparticles with dendritic amine groups on the surface and place them in a conical flask filled with 12ml of 100mM phosphate buffer solution at pH=8, under ultrasonic conditions with a frequency of 40KHz and a power of 240w Ultrasound in ice bath for 20min, evenly dispersed;

[0048] (2) Weigh 150mg of lipase (based on protein content) and place it in 150ml of 100mM phosphate buffer, stir magnetically for 30min at room temperature to fully dissolve the enzyme to form a homogeneous solution, and pour the dispersed magnetic particles prepared in (1) In the system, shake at room temperature for 40 minutes;

[0049] (3) Add 150ml of ethanol dropwise to (2), then stir at 4°C for 30min;

[0050] (4) Add 5ml of 25% glutaraldehyde to (3), shake at room temperature for 8 hours, take the solid after magnetic separation, wash thoroughly with distilled water, and vacuum freeze-dry for 36 hours to obtain magnetically cross-linked lipase a...

Embodiment 3

[0053] (1) Weigh 10mg of functionalized magnetic nanoparticles with dendritic amine groups on the surface and place them in a conical flask filled with 10ml of 100mM phosphate buffer solution at pH=8, under ultrasonic conditions with a frequency of 40KHz and a power of 240w Sonicate in an ice bath for 30 minutes to disperse evenly;

[0054] (2) Weigh 120mg of lipase (based on protein content) and place it in 120ml of 100mM phosphate buffer, stir magnetically for 30min at room temperature to fully dissolve the enzyme to form a homogeneous solution, and pour the dispersed magnetic particles prepared in (1) In the system, shake at room temperature for 40 minutes;

[0055] (3) Add 150ml of acetonitrile dropwise to (2), then stir at 4°C for 30min;

[0056] (4) Add 4ml of 25% glutaraldehyde to (3), shake at room temperature for 8 hours, after magnetic separation, take the solid, wash thoroughly with distilled water, freeze-dry in vacuum for 36 hours, and obtain magnetically cross-l...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
particle diameteraaaaaaaaaa
concentrationaaaaaaaaaa
Login to View More

Abstract

The invention discloses a magnetic crosslinking lipase aggregation as well as a preparation method and application thereof. The magnetism crosslinking lipase aggregation is prepared from functional group magnetic nanoparticles with dendritic polyamine on the surface. The preparation method integrates a crosslinking enzyme aggregation technology and a magnetic immobilized enzyme technology, not only are the enzyme immobilization efficiency and enzyme activity recovery rate of the magnetism immobilized enzyme improved, but also the problem of the poor operational stability of the crosslinking enzyme aggregation is also greatly improved. The prepared magnetic crosslinking lipase aggregation has high activity and excellent operational stability, and can be used as a biocatalyst widely.

Description

technical field [0001] The invention belongs to the field of cross-linked enzyme aggregates, and in particular relates to a magnetic cross-linked lipase aggregate prepared by using functionalized magnetic nanoparticles with dendritic amine groups on the surface, and a preparation method and application thereof. Background technique [0002] As a catalyst, biological enzymes are widely used in industries such as medicine, food, spices, oils, cosmetics and pesticides because of their high efficiency, specificity, mild reaction conditions, and environmental friendliness (Reetz, M.T., 2002. Lipases as practical biocatalysts. Curr. Opin. Chem. Biol. 6, 145–150). Among them, lipase has a wider application prospect because of its wide range of substrates. However, natural lipase has poor operational stability, is easy to inactivate, is not easy to reuse, and is difficult to realize process continuity and automation. After the reaction, it is mixed into the product, which often mak...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/14C12N11/08
Inventor 刘春朝刘英郭晨
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI