Preparation method of silk nano globules coating enzyme and application thereof

A technology of nano-balls and silk, applied in the field of enzyme encapsulation, can solve the problems of unfavorable use in animals, poor biocompatibility, etc., and achieve the effect of protecting storage stability, protecting enzyme activity, and ensuring effective output rate

Active Publication Date: 2017-06-13
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the existing enzyme-encapsulating materials are polymer materials or new inorganic materials, which have poor biocompatibility and are not conducive to use in animals.

Method used

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  • Preparation method of silk nano globules coating enzyme and application thereof
  • Preparation method of silk nano globules coating enzyme and application thereof
  • Preparation method of silk nano globules coating enzyme and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Preparation of Enzyme-coated Silk Nanospheres

[0038] A method for preparing enzyme-encapsulated silk nanospheres, comprising the following steps:

[0039] S1, prepare silk solution, specifically:

[0040] S11, shred silkworm cocoons and place them in 5% NaHCO 3 Boil in the solution, and stir with a glass rod for 30min, repeat this step twice;

[0041] S12. Wash the silk obtained in step S11 with 60°C deionized water for 20 minutes, and repeat this step three times;

[0042] S13, drying the washed silk in an oven at 60°C, then dissolving it in a 9.3M LiBr solution, and dissolving at 60°C for 4 hours;

[0043] S14, dialyze the silk solution obtained in step S13 with deionized water for two days, and change the water every 2 hours;

[0044] S15. Dilute the silk solution obtained in step S14 to 6% w / w;

[0045] S2, encapsulating the enzyme into silk to form silk nanospheres encapsulating the enzyme, specifically:

[0046] S21. Take 1ml of the silk solution...

Embodiment 2

[0053] Example 2 Determination of the activity of enzymes wrapped in silk nanospheres

[0054] (1) Thermostability and storage stability of the enzyme

[0055] By measuring the relative enzyme activity of the enzyme without silk wrapping and the enzyme wrapped with silk at different time points, the thermostability and storage stability of the enzyme before and after wrapping were compared. The result is as figure 2 shown.

[0056] figure 2a represents the relative enzymatic activity of glucose oxidase without silk wrapping and silk wrapping glucose oxidase at 60°C. It can be seen from the figure that at 60°C, glucose oxidase without silk wrapping loses half of its activity in 1 hour; while glucose oxidase wrapped in silk loses half of its activity after 30 hours; The thermal stability of the enzyme encapsulated by nanospheres is effectively protected.

[0057] figure 2 b represents the storage stability of horseradish peroxidase without silk wrapping and silk wrappin...

Embodiment 3

[0067] Embodiment 3 in the application of antialcoholic medicine

[0068] Alcohol oxidase (AOx) and catalase (Cat) are wrapped in silk nanospheres to prepare a protease-resistant anti-alcoholic drug, and its effect is as follows: Image 6 shown.

[0069] In the simulated intestinal fluid, add the alcohol enzyme without silk nanospheres or the alcoholase with silk nanospheres, and compare the relative enzyme activities of the two, as shown in Image 6 As shown in a, the alcohol enzyme wrapped with silk nanospheres still retains more than 90% of its activity after 2 hours, while the relative activity of the alcoholase without silk nanospheres wrapped rapidly drops to 5% after 20 minutes. This is because the drug containing protease reaches the intestinal juice after oral administration. The protease in the intestinal juice will promote the hydrolysis of protein, and the essence of most enzymes is protein, which leads to the rapid decomposition and inactivation of the enzyme in ...

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Abstract

The invention discloses a preparation method of silk nano globules coating an enzyme. The preparation method comprises the following steps of: S1: preparing a silk solution; and S2: wrapping the enzyme into the silk to form the silk nano globules coating the enzyme. The prepared silk nano globules can be applied to the medical field, and in particular to an antialcoholic drug. The silk nano globules prepared by the method can effectively protect the activity of the wrapped enzyme to realize cascade reactions of various enzymes, and meanwhile have good biocompatibility.

Description

technical field [0001] The invention relates to the technical field of enzyme encapsulation, in particular to a method for preparing enzyme encapsulating silk nano balls and an application thereof. Background technique [0002] Enzymes are essential natural catalysts in living organisms, and are closely related to almost all biological reactions, such as protein synthesis and metabolism. However, natural biological enzymes are very fragile, easily lose their activity when exposed to normal temperature, and have high requirements on the reaction environment, so it is very important to protect the activity of enzymes such as thermal stability and storage stability. [0003] In addition, most of the biological reactions are completed through the series reactions of various enzymes, and intermediate products will be produced during these series reactions, and some intermediate products are mostly harmful to the human body. Therefore, researchers have simulated the biological ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/51A61K38/44A61K47/46A61P39/02
CPCA61K9/5176A61K38/44A61K38/443C12Y101/03004C12Y111/01006C12Y111/01007A61K2300/00
Inventor 陈政维刘向阳林友辉
Owner XIAMEN UNIV
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