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A kind of hydrogel containing lipase-inorganic hybrid nanoflowers and its preparation method and application

A lipase and nanoflower technology, applied in biochemical equipment and methods, immobilized on or in inorganic carriers, nanotechnology, etc., can solve problems such as reduced stability, low mechanical strength, and enzyme diffusion limitations, and achieve stable Improved performance, improved catalytic activity, and avoided mass transfer obstructions

Active Publication Date: 2020-11-06
DALIAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the mechanical strength of this nanoflower-shaped catalytic material is very low, and it is easily affected by the external environment or external forces, resulting in reduced stability, and requires centrifugation to separate the nanoflowers from the substrate, which is not conducive to rapid industrial production.
[0004] At the same time, many domestic and foreign scholars have immobilized enzymes into polymer hydrogels with better biocompatibility, better thermal stability and porous through embedding methods, such as: sodium alginate gel microspheres, graphene Polymer composite hydrogels, chitosan-clay nanocomposite membranes, silicon polymer gel microspheres, etc., have improved the mechanical properties of enzyme-catalyzed materials and improved the stability of enzymes. Unfortunately, after immobilization The diffusion of the enzyme is greatly restricted, and at the same time, a small amount of the enzyme is dissolved from the polymer gel, resulting in a serious loss of enzyme activity

Method used

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  • A kind of hydrogel containing lipase-inorganic hybrid nanoflowers and its preparation method and application
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  • A kind of hydrogel containing lipase-inorganic hybrid nanoflowers and its preparation method and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] (1) Use PBS to prepare a lipase solution with an initial enzyme concentration of 0.1 mg / mL, take 3 mL of the lipase solution, add 20 μL of a copper chloride solution with a concentration of 120 mM, shake well, and incubate at room temperature (25°C) After 48 hours, the precipitate was collected, washed three times with deionized water, and centrifuged at 4°C and 3500 rpm for 5 minutes with a low-temperature high-speed centrifuge, and the precipitate was freeze-dried for 24 hours to obtain a blue powder that is lipase-inorganic hybrid nano flower;

[0049] (2) Get 3g of 15% polyvinyl alcohol (PVA) PBS solution, then add 1g of 5% chitosan solution utilizing 2% aqueous acetic acid configuration, mix well; add the fat prepared in (1) in the mixed solution Enzyme-inorganic hybrid nanoflowers, added in an amount of 1 wt% of the mass of the mixed solution, stirred evenly at 25°C;

[0050] (3) The uniformly mixed solution is subjected to cyclic freezing-thawing, specifically: ...

Embodiment 2

[0060] (1) Use PBS to prepare a lipase solution with an initial enzyme concentration of 0.1 mg / mL, take 3 mL of the lipase solution, add 20 μL of a copper chloride solution with a concentration of 120 mM, shake well, and incubate at room temperature (25°C) After 48 hours, collect the precipitate, wash the precipitate three times with deionized water, and centrifuge at 4°C and 3500 rpm for 5 minutes with a low-temperature high-speed centrifuge, freeze-dry the precipitate for 24 hours, and obtain a blue powder that is lipase-inorganic hybrid nanoflowers;

[0061] (2) Get 3g of 15% polyvinyl alcohol (PVA) PBS solution, then add 1g of 5% chitosan solution utilizing 2% aqueous acetic acid configuration, mix well; add the fat prepared in (1) in the mixed solution Enzyme-inorganic hybrid nanoflowers, added in an amount of 2wt% of the mass of the mixed solution, stirred evenly at 25°C;

[0062] (3) The uniformly mixed solution was subjected to cycle freezing-thawing, freezing at -20°...

Embodiment 3

[0064] (1) Use PBS to prepare a lipase solution with an initial enzyme concentration of 0.1 mg / mL, take 3 mL of the lipase solution, add 20 μL of a copper chloride solution with a concentration of 120 mM, shake well, and incubate at room temperature (25°C) After 48 hours, collect the precipitate, wash the precipitate three times with deionized water, and centrifuge at 4°C and 3500 rpm for 5 minutes with a low-temperature high-speed centrifuge, freeze-dry the precipitate for 24 hours, and obtain a blue powder that is lipase-inorganic hybrid nanoflowers;

[0065] (2) Get 3g of 15% polyvinyl alcohol (PVA) PBS solution, then add 1g of 5% chitosan solution utilizing 2% aqueous acetic acid configuration, mix well; add the fat prepared in (1) in the mixed solution Enzyme-inorganic hybrid nanoflowers, added in an amount of 3 wt% of the mass of the mixed solution, stirred evenly at 25°C;

[0066] (3) The uniformly mixed solution was subjected to cycle freezing-thawing, freezing at -20...

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Abstract

The invention discloses a hydrogel-type enzyme catalytic material synthesized via secondary immobilization and containing lipase and inorganic hybrid nanometer flowers and a preparation method and application of the hydrogel-type enzyme catalytic material. Lipase is taken as an organic component and is used to synthesize lipase and inorganic hybrid nanometer flowers together with inorganic metal ions in a self-assembly manner; the obtained nanometer flowers are embedded into a gel carrier system having relatively good biocompatibility and containing at least two hydrophilic polymer; and a repeated freezing-unfreezing method is adopted to obtain hydrogel containing lipase and inorganic hybrid nanometer flowers. The hydrogel has a unique tridimensional network to stabilize an enzyme in a catalytic system, thereby achieving secondary immobilization. Through secondary immobilization, lipase is protected from being affected by the external environment, so that the stability and reutilization of the enzyme are improved. The catalytic material doesn't have to be separated from a reactant, so that the step of separating the enzyme from a substrate is omitted. The material has bright application prospects in the field of enzyme immobilization.

Description

technical field [0001] The invention belongs to the field of enzyme immobilization, and relates to a hydrogel enzyme catalytic material containing lipase-inorganic hybrid nanoflower synthesized by secondary immobilization, a preparation method and application thereof. Background technique [0002] Lipase is a kind of enzyme that can catalyze the hydrolysis reaction, esterification reaction and transesterification reaction of glyceride in micro-aqueous or non-aqueous phase system. Because of its good water solubility, reaction specificity and high catalytic efficiency , biocompatibility, low toxicity and other advantages make it have certain applications in energy, food, washing, leather, pharmaceutical, textile and paper making industries, so it has been widely concerned and researched by scholars at home and abroad. However, there are still many defects in the practical application of lipase as an industrial catalyst. It is prone to decrease in stability, loss of enzyme act...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/14C12N11/084C12N11/10B82Y40/00B82Y30/00A23L27/20
CPCA23L27/206B82Y30/00B82Y40/00C12N9/20C12N11/08C12N11/10C12N11/14C12Y301/01003
Inventor 田晶井木子费旭李尧任伟帆赵丽媛徐龙权王一
Owner DALIAN POLYTECHNIC UNIVERSITY