Preparation method of magnetic chitosan compound microsphere immobilized marine alkaline proteinase

A technology of composite microspheres and chitosan, applied in the direction of being fixed on or in an inorganic carrier, immobilized on or in an organic carrier, etc., can solve the problems of application difficulty, not, and the difficulty of free enzyme to meet user requirements, etc.

Inactive Publication Date: 2012-06-27
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to certain problems in the cost, high efficiency and stability of free enzymes, its application still faces many difficulties.
Therefore, for modern industry, it is difficult for free enzymes to meet user requirements in use, and it is not an ideal enzyme.

Method used

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  • Preparation method of magnetic chitosan compound microsphere immobilized marine alkaline proteinase
  • Preparation method of magnetic chitosan compound microsphere immobilized marine alkaline proteinase
  • Preparation method of magnetic chitosan compound microsphere immobilized marine alkaline proteinase

Examples

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Effect test

example 1

[0054] Put 30mL 0.5mol / L FeCl 2 Solution and 50mL 0.5mol / L FeCl 3 The solution is evenly mixed, and 8.0g PEG4000 is added. After it is fully dissolved, it is quickly added dropwise to 100mL of 5% diluted ammonia water under stirring, and the pH value of the solution is adjusted to 9.0 with ammonia water. After stirring for 30 minutes, ultrasonic for 10 minutes, 80°C water bath maturation for 30 minutes, and ultrasonic for 10 minutes to make the solution system uniform and fine. Finally, the supernatant liquid is poured under the adsorption of the magnet, washed with distilled water to neutrality, and freeze-dried in vacuum to obtain Fe 3 O 4 Magnetic core.

[0055] Accurately weigh 0.5g of chitosan, dissolve it in 20mL of 5wt% acetic acid solution to obtain 2.5wt% of chitosan acetic acid solution, add 0.5g Fe into the system 3 O 4 Magnetic core and mix well. Add dropwise to a mixed system consisting of 5mL of emulsifier Span-80 and 80mL of liquid paraffin with slow stirring. Afte...

example 2

[0058] Put 25mL 0.6mol / L FeCl 2 Solution and 40mL 0.6mol / L FeCl 3 The solution is evenly mixed, and 6.0 g of PEG4000 is added. After it is fully dissolved, it is quickly added dropwise to 120 mL of 5 wt% dilute ammonia with stirring, and the pH of the solution is adjusted to 9.0 with ammonia. After stirring for 25 minutes, ultrasonic for 15 minutes, 70°C water bath maturation for 40 minutes, and ultrasonic for 20 minutes to make the solution system uniform and fine. Finally, the supernatant liquid is poured under the adsorption of the magnet, washed with distilled water to neutrality, and freeze-dried in vacuum to obtain Fe 3 O 4 Magnetic core.

[0059] Accurately weigh 0.5g of chitosan and dissolve it in 20mL of 5wt% acetic acid solution to obtain 2.5wt% of chitosan acetic acid solution. Add the above 0.25g Fe to this system 3 O 4 Magnetic core and mix well. Add dropwise to a mixed system consisting of 5mL of emulsifier Span-80 and 80mL of liquid paraffin under slow stirring. Af...

example 3

[0062] Put 40mL 0.4mol / L FeCl 2 Solution and 60mL 0.4mol / L FeCl 3 The solution is evenly mixed, 10.0g PEG4000 is added, and after it is fully dissolved, it is quickly added dropwise to 80mL of 5wt% dilute ammonia water under stirring, and the pH value of the solution is adjusted to 9.0 with ammonia water. After stirring for 40 minutes, ultrasonic for 10 minutes, aging in a water bath at 90°C for 20 minutes, and ultrasonic for 20 minutes to make the solution system uniform and fine. Finally, the supernatant liquid is poured under the adsorption of the magnet, washed with distilled water to neutrality, and freeze-dried in vacuum to obtain Fe 3 O 4 Magnetic core.

[0063] Accurately weigh 0.8g chitosan and dissolve it in 20mL 5wt% acetic acid solution to obtain 2.5wt% chitosan acetic acid solution. Add 0.5g Fe into the system 3 O 4 Magnetic core and mix well. Add dropwise to a mixed system consisting of 5mL of emulsifier Span-80 and 80mL of liquid paraffin with slow stirring. After ...

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Abstract

The invention relates to a preparation method of magnetic chitosan compound microsphere immobilized marine alkaline proteinase 894. The magnetic chitosan compound microsphere immobilized marine alkaline proteinase is prepared from Fe3O4 magnetic chitosan compound microsphere and magnetic chitosan compound microsphere immobilized marine alkaline proteinase 894. The method further improves the using efficiency, lowers using and production cost, obtains the immobilized enzyme with stable usability, high activity recovery and long using period and has wide application on food, textile and feed.

Description

Background technique [0001] The present invention relates to the field of marine microbial enzymes, in particular to a preparation method of immobilized marine alkaline protease 894 with magnetic chitosan composite microspheres. [0002] Alkaline protease is an important type of industrial enzyme, mainly derived from alkaliphilic microorganisms. It can catalyze the hydrolysis of protein into small peptides and amino acids. It is widely used in washing, food, leather and silk industries, and is used in medicine, environmental protection, chemical industry and basic research. China also shows a broad application prospect. Marine alkaline protease 894 is a new type of marine enzyme developed by our laboratory using the metabolites of marine microorganisms in our country through modern bioengineering methods. Studies have shown that the enzyme has high activity, stable storage, and good compatibility. It is widely used in textiles and feed. [0003] In order to adapt to the life limit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/14C12N11/10C12N11/04
Inventor 王海英孙谧牛瑞
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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