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Silica-gel-carrier surface macroporous chitin substrate for lysozyme affinity chromatography

A chitin and lysozyme technology, applied in the directions of enzymes, enzymes, biochemical equipment and methods, can solve the problems of difficulty in extracting surfactants and porogens, and achieve excellent surface porous structure and small non-specific adsorption. Effect

Inactive Publication Date: 2008-05-14
QINGDAO UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method usually needs to use a large amount of organic solvents such as liquid paraffin, and it is difficult to extract the surfactant and porogen contained in the microbeads.

Method used

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  • Silica-gel-carrier surface macroporous chitin substrate for lysozyme affinity chromatography
  • Silica-gel-carrier surface macroporous chitin substrate for lysozyme affinity chromatography

Examples

Experimental program
Comparison scheme
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Embodiment Construction

[0028] Add 2g of refined chitosan and 10g of porogen polyethylene glycol to 1mol / L 100mL acetic acid solution, stir magnetically for 30min, remove insoluble residue after filtering through filter cloth; add inorganic silane reagent γ-glycidoxypropylene to the filtrate 2mL of trimethoxysilane, magnetically stirred for 30min, then added 50g of 60-80 mesh silica gel to the solution while stirring, stirred evenly, and ultrasonicated for 40min; poured the suspension into a petri dish, stirred evenly, stood at room temperature for 48h, and baked at 40°C Dry. Place the obtained solid in 0.5mol / L 100mL ammonia solution, extract at 80°C, replace the ammonia solution every 2h, take out the solid after 8h, wash with distilled water until neutral, dry at 40°C, place in 95mL methanol, add 5mL acetic acid anhydride, stir for 2 hours, wash the solid with distilled water until it is neutral, place it in 0.1mol / L hydrochloric acid, stir to suspend it, slowly add sodium nitrite solution, stir f...

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PUM

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Abstract

The invention opened a new macro porous silica gel loading surface Chitin base for the lysozym affinity chromatography. The invention is to load the chitosan in the silica gel firstly, the cross linker is the gamma-epoxypropyl trimethoxy silane with the epoxy groups and the silane. After acetylating, the product would be get. The grain size of the base can be adjusted by adjusting the size of the silica gel carrier. The polyethylene glycol 20000 is to make the hole to fit for the protein. The nonspecific base is small because of the free amino group is removed by the hydrocarbonylation. The product can be used for the purifying of the lysozym in the egg white.

Description

【Technical field】 [0001] The invention relates to a lysozyme affinity chromatography substrate, in particular to a silica gel-loaded chitin substrate with a surface macropore structure suitable for the lysozyme affinity chromatography. 【Background technique】 [0002] Lysozyme (Lysozyme, EC 3.2.1.17), also known as mureinase, or N-acetylmurinase, is a hydrolytic enzyme that specifically acts on the cell wall of microorganisms and is widely found in animals, plants and microorganisms In it, it has the function of sterilizing and breaking the wall. Lysozyme has stable chemical properties and good biocompatibility, is safe and non-toxic to the human body, and has no side effects. It has been widely used in clinical treatment and diagnosis such as coagulation, decomposition of pus and tissue repair. With the development of bioengineering, lysozyme has become an essential tool enzyme for plasmid, inclusion body and cell wall breaking in genetic engineering and cell engineering. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/36
Inventor 李峰牛淑妍张书圣
Owner QINGDAO UNIV OF SCI & TECH
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