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Method for immobilizing trypsinase by chitosan microspheres

A technology of chitosan microspheres and trypsin, which is applied in the field of chitosan microspheres immobilized trypsin and chitosan immobilized enzymes, can solve the problems of high impurities and low activity of products, and achieve high recovery rate of enzyme activity , high vitality, and good operational stability

Inactive Publication Date: 2010-07-14
WUHAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As mentioned in the patent publication No. CN101285060A, most of the previous studies have some defects, such as the obtained product has many impurities and low activity.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] (1) Preparation of chitosan microspheres: a certain amount of chitosan was weighed and dissolved in an acetic acid solution with a mass concentration of 1% to prepare a chitosan solution with a mass concentration of 2%. Add liquid paraffin, span-80 and porogen ethyl acetate into the container, the amount of span-80 is 0.5% of the paraffin mass, and the porogen ethyl acetate is 0.01% of the paraffin mass; add slowly under high-speed stirring Chitosan solution, the chitosan solution / paraffin mass ratio is 1:1.5, keep stirring at high speed for 40 minutes, so that the chitosan droplets are evenly dispersed to obtain a suspended emulsion; then slowly add to the emulsion while stirring Glutaraldehyde solution, so that the mass concentration of glutaraldehyde in the emulsion reaches 1%, and react at 45° C. for 1 hour. After standing and cooling, the obtained emulsion is poured into an equal volume of NaOH / ethanol mixed solution, fully mixed and oscillated, then left to stand,...

Embodiment 2

[0017] (1) Preparation of chitosan microspheres: a certain amount of chitosan was weighed and dissolved in an acetic acid solution with a mass concentration of 2%, to prepare a chitosan solution with a mass concentration of 3%. Add liquid paraffin, span-80 and porogen ethyl acetate into the container, the amount of span-80 is 0.1% of the paraffin mass, and the porogen ethyl acetate is 0.05% of the paraffin mass; add slowly under high-speed stirring Chitosan solution, the mass ratio of chitosan solution / paraffin wax is 1:2, keep stirring at high speed for 60 minutes, so that the chitosan droplets are evenly dispersed to obtain a suspended emulsion; then slowly add to the emulsion while stirring Glutaraldehyde solution, so that the mass concentration of glutaraldehyde in the emulsion reaches 6%, and react at 4° C. for 10 h. After standing and cooling, the obtained emulsion is poured into an equal volume of NaOH / ethanol mixed solution, fully mixed and oscillated, then left to sta...

Embodiment 3

[0020] (1) Preparation of chitosan microspheres: a certain amount of chitosan was weighed and dissolved in an acetic acid solution with a mass concentration of 3% to prepare a chitosan solution with a mass concentration of 4%. Add liquid paraffin, span-80 and porogen ethyl acetate into the container, the amount of span-80 is 1% of the paraffin mass, and the porogen ethyl acetate is 0.1% of the paraffin mass; add slowly under high-speed stirring Chitosan solution, the chitosan solution / paraffin mass ratio is 1:3, keep stirring at high speed for 100min, so that the chitosan droplets are evenly dispersed to obtain a suspended emulsion; then slowly add to the emulsion while stirring Glutaraldehyde solution, so that the mass concentration of glutaraldehyde in the emulsion reaches 3%, and react at 30° C. for 4 hours. After standing and cooling, the obtained emulsion is poured into an equal volume of NaOH / ethanol mixed solution, fully mixed and oscillated, then left to stand, discard...

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Abstract

The invention provides a method for immobilizing trypsinase by chitosan microspheres, which belongs to the technical field of food processing. The invention uses chitosan as immobilizing carriers, and uses a reverse-phase suspension method for preparing the chitosan microspheres after the chitosan is dissolved in an acetic acid solution, then, glutaric dialdehyde is used as a cross linking agent for cross linking activation on the chitosan microspheres, the chitosan microspheres after the cross linking activation are mixed with a trypsinase solution for immobilization, and the immobilized trypsinase is obtained. The trypsinase is one of the enzyme preparations in most common use of protein modification and hydrolysis in food industry. The invention uses the chitosan for immobilizing the trypsinase, so the trypsinase can be used repeatedly, the utilization efficiency of the trypsinase is improved, the enzyme consumption in the protein hydrolysis process and the production cost are reduced, and the invention can be widely used for the hydrolysis process and the modification of protein such as cow milk, meat, aquatic products and the like.

Description

technical field [0001] The invention relates to a method for immobilizing enzymes with chitosan, in particular to a method for immobilizing trypsin with chitosan microspheres, and belongs to the technical field of food processing. Background technique [0002] Trypsin is a protease commonly used in the food industry, such as the modification of milk protein, the enzymatic hydrolysis of fish, shrimp, and animal protein to recover protein. However, enzyme preparations are generally expensive, and enzymes are easily inactivated during use and are usually not recyclable, which often leads to increased production costs; in addition, due to the non-recyclability of enzymes, it also leads to the risk of introducing foreign substances into food. The immobilized enzyme can improve the stability of the enzyme, maintain its activity for a long time, and can realize recycling after immobilization, which greatly reduces the cost of enzyme use, so immobilized trypsin has very important pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/10
Inventor 夏文水谭丽姜启兴
Owner WUHAN POLYTECHNIC UNIVERSITY
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