Hydrogel containing protease-inorganic hybrid nanoflower, and preparation method and applications thereof

A technology of protease and nanoflower, which is applied in the field of hydrogel-type enzyme catalytic materials and its preparation, can solve the problems of reduced stability, low mechanical strength, and restriction of enzyme diffusion, so as to achieve improved stability, improved catalytic activity, and avoid transmission. The effect of mass hindrance

Active Publication Date: 2018-06-08
山东同益光刻胶材料科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 throu...

Method used

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  • Hydrogel containing protease-inorganic hybrid nanoflower, and preparation method and applications thereof
  • Hydrogel containing protease-inorganic hybrid nanoflower, and preparation method and applications thereof
  • Hydrogel containing protease-inorganic hybrid nanoflower, and preparation method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1～16

[0053] (1) Use PBS to prepare papain solutions with initial enzyme concentrations of 0.1mg / mL, 0.4mg / mL, 0.8mg / mL, and 1.2mg / mL, respectively, take 6mL of the papain solution, and add a concentration of 120mM chloride 40 μL of copper solution, shake well and let stand at room temperature (25°C) for 72 hours, collect the precipitate, wash the precipitate three times with deionized water, and centrifuge at 4°C, 3500rpm for 5min with a low-temperature high-speed centrifuge, and the precipitated part Freeze-dry for 24 hours to obtain a blue powder, and obtain papain-inorganic hybrid nanoflowers with initial enzyme concentrations of 0.1mg / mL, 0.4mg / mL, 0.8mg / mL, and 1.2mg / mL;

[0054] (2) Dissolve a certain amount of polyvinyl alcohol and chitosan in a certain amount of phosphate buffer, mix uniformly to obtain a gel carrier system, add a certain amount of papain-inorganic hybrid nanoflowers of step (1) , dissolved, and mixed uniformly; the mass percentages of papain-inorganic hybr...

Embodiment 17

[0068] Under different pH values, measure the enzyme activity of papain-inorganic hybrid nanoflower hydrogel containing papain of the present invention, specific method is: weigh papain-inorganic hybrid nanoflower hydrogel ( The enzyme content in the hydrogel is equal to the enzyme content in the nanoflower), and it was dispersed in 2mL PBS (0.01mol / L, and the pH value was adjusted to 6.5, 7.5, 8.0 and 9.0 with NaOH, respectively, to prepare Hydrogel liquid. According to the above-mentioned protease enzyme activity assay method, each reagent is added in the hydrogel liquid, after mixing, shock and react for 5min, measure the absorbance value at 253nm, and calculate the enzyme activity. The results are shown in Tables 2 to 5. As a comparison , the enzyme activity of papain-inorganic hybrid nanoflower (0.4 mg enzyme equivalent) was measured under the same pH condition, and the results are shown in Table 6.

[0069] Table 2. Enzyme activity of papain-inorganic hybrid nanoflower h...

Embodiment 18

[0082] Determination of the protein degradation performance of papain-inorganic hybrid nanoflower hydrogels with different nanoflower contents, specifically: Weigh 2 g of papain-inorganic hybrid nanoflower hydrogels with different nanoflower contents prepared in Examples 13-16 glue, add it to 20g of white wine, and shake it at room temperature for 24 hours. Proteins and amino acids in the wine were then detected.

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Abstract

The invention discloses a hydrogel type enzyme catalytic material which contains protease-inorganic hybrid nanoflower and is prepared through secondary immobilization synthesis, and a preparation method and applications thereof. According to the preparation method, protease is taken as an organic component, and is subjected to self-assembling with inorganic metal ions so as to obtain the protease-inorganic hybrid nanoflower; the protease-inorganic hybrid nanoflower is embedded with a gel carrier system which is excellent in biocompatibility and contains at least two hydrophilic polymers, repeat freezing-unfreezing method is adopted to obtain the hydrogel containing protease-inorganic hybrid nanoflower. The unique tridimensional network of the hydrogel containing protease-inorganic hybrid nanoflower is capable of realizing stable existing of enzymes in a catalytic system, and secondary immobilization is achieved. According to the preparation method, secondary immobilization is adopted,so that protease is protected from external environment influences, enzyme stability is improved, enzyme reusability is improved. Separation of the catalytic material with reactants is not necessary,so that a step of separation of enzymes with substrates is avoided, and the application prospect in the field of enzyme immobilization is promising.

Description

technical field [0001] The invention belongs to the field of enzyme immobilization, and relates to a hydrogel-type enzyme catalytic material containing protease-inorganic hybrid nanoflower synthesized by secondary immobilization, a preparation method and application thereof. Background technique [0002] As an enzyme that catalyzes protein hydrolysis, protease has been widely used in biochemistry, biomedicine, food, textile, chemical industry and other fields. However, since most proteases are easily destroyed under the environmental conditions of strong acid, strong alkali, high temperature and heavy metal ions, the production and application of enzymes are greatly restricted. Therefore, how to improve the stability of enzymes and expand the utilization range of enzymes has become a research hotspot. [0003] In recent years, scholars at home and abroad have made breakthroughs in the synthesis of organic-inorganic hybrid nanoflowers with three-dimensional flower-shaped str...

Claims

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Application Information

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IPC IPC(8): C12N9/50B01J13/00C12H1/02
CPCB01J13/0065C12H1/02C12N9/50
Inventor 费旭支慧田晶李尧张海洋赵丽媛徐龙权王一
Owner 山东同益光刻胶材料科技有限公司
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