Preparation method of magnetic chitosan compound microsphere immobilized marine alkaline proteinase

A technology of composite microspheres and chitosan, which is applied in the direction of being immobilized on or in an inorganic carrier, immobilized on or in an organic carrier, etc., can solve the problems that the free enzyme is difficult to meet the requirements of users, difficult to apply, not to be used, etc.

Inactive Publication Date: 2010-11-24
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to certain problems in the cost, high efficiency and stability of free enzymes, its application still faces many difficul

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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example 1

[0054] 30mL 0.5mol / L FeCl 2 solution and 50mL of 0.5mol / L FeCl 3 Mix the solution evenly, add 8.0g PEG4000, after fully dissolved, quickly drop it into 100mL 5% dilute ammonia water under stirring, and continue to adjust the pH value of the solution to 9.0 with ammonia water. After continuing to stir for 30 minutes, sonicate for 10 minutes, mature in an 80°C water bath for 30 minutes, and sonicate again for 10 minutes to make the solution system uniform and fine. Finally, under the adsorption of the magnet, the supernatant was decanted, washed with distilled water until neutral, and vacuum freeze-dried to obtain Fe 3 o 4 magnetic core.

[0055] Accurately weigh 0.5g chitosan, dissolve in 20mL 5wt% acetic acid solution to obtain 2.5wt% chitosan acetic acid solution, add 0.5g Fe 3 o 4 Nucleate and mix well. Slowly stirred into the mixed system consisting of 5mL of emulsifier Span-80 and 80mL of liquid paraffin drop by drop. After electric stirring for 10 minutes, quickly d...

example 2

[0058] 25mL 0.6mol / L FeCl 2 solution and 40mL of 0.6mol / L FeCl 3The solution was mixed evenly, and 6.0 g of PEG4000 was added, and after being fully dissolved, it was rapidly added dropwise into 120 mL of 5 wt % dilute ammonia water under stirring, and the pH value of the solution was adjusted to 9.0 with ammonia water. After continuing to stir for 25 minutes, sonicate for 15 minutes, mature in a water bath at 70°C for 40 minutes, and sonicate again for 20 minutes to make the solution system uniform and fine. Finally, the supernatant was decanted under the adsorption of a magnet, washed with distilled water until neutral, and vacuum freeze-dried to obtain Fe 3 o 4 magnetic core.

[0059] Accurately weigh 0.5g chitosan, dissolve in 20mL 5wt% acetic acid solution to obtain 2.5wt% chitosan acetic acid solution, add above-mentioned 0.25g Fe in this system 3 o 4 Nucleate and mix well. Slowly stirred into the mixed system consisting of 5mL of emulsifier Span-80 and 80mL of liq...

example 3

[0062] 40mL 0.4mol / L FeCl 2 solution and 60mL of 0.4mol / L FeCl 3 Mix the solution evenly, add 10.0g PEG4000, after fully dissolved, quickly drop it into 80mL 5wt% dilute ammonia water under stirring, and continue to adjust the pH value of the solution to 9.0 with ammonia water. After continuing to stir for 40 minutes, sonicate for 10 minutes, mature in a 90°C water bath for 20 minutes, and sonicate again for 20 minutes to make the solution system uniform and fine. Finally, the supernatant was decanted under the adsorption of a magnet, washed with distilled water until neutral, and vacuum freeze-dried to obtain Fe 3 o 4 magnetic core.

[0063] Accurately weigh 0.8g chitosan, dissolve in 20mL 5wt% acetic acid solution to obtain 2.5wt% chitosan acetic acid solution, add 0.5g Fe 3 o 4 Nucleate and mix well. Slowly stirred into the mixed system consisting of 5mL of emulsifier Span-80 and 80mL of liquid paraffin, and then quickly added 5mL of 10wt% glutaraldehyde solution drop...

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Abstract

The invention relates to a preparation method of magnetic chitosan compound microsphere immobilized marine alkaline proteinase. 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 invention relates to the field of marine microbial enzymes, in particular to a method for preparing marine alkaline protease 894 immobilized by magnetic chitosan composite microspheres. [0002] Alkaline protease is an important class of industrial enzymes, mainly derived from alkaliphilic microorganisms, which can catalyze the hydrolysis of proteins into small peptides and amino acids. It is widely used in washing, food, leather and silk industries. It also shows broad application prospects. 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 used in food, It is widely used in textile and feed. [0003] In order to adapt to the life-limiting environment of the ocean, marine microbial enzymes exhibit more uniqu...

Claims

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Application Information

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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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