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Preparation method of chitosan microspheres for cell immobilization and drug delivery

A technology of chitosan microspheres and cell immobilization, applied in the direction of microorganism-based methods, microsphere preparation, biochemical equipment and methods, etc., can solve the problems of uneven size, time-consuming and labor-intensive, unsuitable for industrial production, etc. Achieve the effects of increasing deacetylation speed, increasing production capacity, cell viability retention and reusability

Pending Publication Date: 2021-10-29
上海信卓实业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, chitosan microspheres prepared by manual knocking and spray drying are not uniform in size, and the preparation process is time-consuming and labor-intensive, which is not suitable for industrial production.

Method used

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  • Preparation method of chitosan microspheres for cell immobilization and drug delivery

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A preparation method for chitosan microspheres for cell immobilization and drug delivery, comprising the following steps:

[0037] 1) Weigh chitin and chitin deacetylase according to a mass ratio of 1000:1, add water to dissolve, place in a microwave device, treat at 50°C, microwave power 600W for 10 minutes, and inactivate the enzyme to obtain a chitosan solution.

[0038] 2) Using tetrasodium glutamic acid diacetate to prepare a chelating agent with a concentration of 0.02mol / L, the chelating agent and chitosan solution are fully stirred according to the molar ratio of 10:0.2, mixed evenly, and the solution is observed until the solution changes from clarification to From cloudy to clear.

[0039] 3) Add 0.5g / L carbonate to the solution in step 2) for sedimentation, stir fully, improve the extraction and removal ability of the chelating agent to residual metal ions, and obtain a crude chitosan solution.

[0040] 4) Add complex biological enzymes (specific keratinase ...

Embodiment 2

[0050] A preparation method for chitosan microspheres for cell immobilization and drug delivery, comprising the following steps:

[0051] 1) Weigh chitin and chitin deacetylase according to a mass ratio of 1000:1, add water to dissolve, place in a microwave device, treat at 50°C, microwave power 400W for 10 minutes, and inactivate the enzyme to obtain a chitosan solution.

[0052] 2) Use citric acid to prepare a chelating agent with a concentration of 0.01mol / L. The chelating agent and chitosan solution are fully stirred according to the molar ratio of 20:1, and mixed evenly. Observe the change of the solution until the solution changes from clear to cloudy and then to clear. .

[0053] 3) Add 2g / L carbonate to the solution in step 2) for sedimentation, stir fully, improve the extraction and removal ability of the chelating agent to residual metal ions, and obtain a crude chitosan solution.

[0054] 4) Add complex biological enzymes (specific keratinase and lipase) to the cru...

Embodiment 3

[0059] Using the immobilized cells prepared in Example 1, the continuous conversion production of niacin was carried out by using a packed bed bioreactor. Nicotinic acid, also known as nicotinic acid and anti-peaceae factor, is converted into nicotinamide in the human body and participates in lipid metabolism in the body, the oxidation process of tissue respiration and the process of anaerobic decomposition of carbohydrates. A packed bed bioreactor is built by using chromatographic columns, peristaltic pumps, super water baths and other related instruments, and the device is used for continuous conversion and production of niacin. The optimum reaction conditions for continuous transformation are: temperature 35°C, pH 7.0, substrate 3-cyanopyridine concentration 350mmol·L -1 , the volume ratio of immobilized cells to substrate 3-cyanopyridine is 1:1, the residence time is 35.2min, and the transformation capacity of cross-linked PVA-CS immobilized cells is 348.4g (3-cyanopyridin...

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Abstract

The invention discloses a preparation method of chitosan microspheres for cell immobilization and drug delivery, and relates to the field of preparation of chitosan microspheres. According to the method, chitin is deacetylated by combining a compound enzyme with a microwave induction process technology, the chitosan is subjected to heavy metal green chelation by adopting tetrasodium glutamate diacetate or citric acid, and the extraction and removal capabilities of a chelating agent on residual metal ions are further improved by further utilizing a carbonate dissolution promoting technology; and trace proteins and lipids in crude chitosan products are hydrolyzed and separated by selecting a proper biological enzyme, refined chitosan is obtained, enzymatic molecular modification is performed on the refined chitosan by using transglycosides or oxidase, and finally the chitosan microspheres are prepared by using a green and environment-friendly spray drying method. The prepared chitosan microspheres are smooth in shape and uniform in size, the particle size is 1-250 microns, the chitosan microspheres are of a core-shell structure, and the application range of chitosan in the fields of medicine, food and other industries is widened by utilizing chitosan immobilized enzyme.

Description

technical field [0001] The invention relates to the field of preparation of chitosan microspheres, in particular to a preparation method of chitosan microspheres for cell immobilization and drug delivery. Background technique [0002] Chitosan (chitosan) is the product of deacetylation of chitin. It is the only alkaline polysaccharide among natural polysaccharides. It is widely distributed in the cell walls of crustaceans, insect shells and fungi, with an annual output of tens of billion tons. As a new type of green polymer material, chitosan contains a large number of coordinating groups such as amino and hydroxyl groups, has high reactivity, and can undergo alkylation, esterification, acylation, carboxylation, hydrolysis, and chelation. Combination and other chemical reactions, introducing a variety of functional groups, changing its physical and chemical properties, can also make chitosan into different application forms such as chitosan microspheres, chitosan nanoparticl...

Claims

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Application Information

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IPC IPC(8): A61K9/16A61K47/36A61K47/32B01J13/02B01J13/04C12N11/10C12N11/084C12P19/26C12R1/645
CPCC12N11/10C12N11/084C12P19/26A61K9/1652A61K9/1635B01J13/02B01J13/043
Inventor 朱萌丁振中方祥管平海高小燕徐俊山
Owner 上海信卓实业有限公司
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