Method for inhibiting activity of amylase

An amylase and activity technology, which is applied in the field of improving the enzyme activity of food and medicine, and achieves the effects of simple method, high yield and uniform distribution

Active Publication Date: 2018-03-30
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the effect of polysaccharide nanoparticles on amylase activity has not been reported

Method used

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  • Method for inhibiting activity of amylase
  • Method for inhibiting activity of amylase
  • Method for inhibiting activity of amylase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] (1)①Preparation of retrograde starch nanoparticles

[0053] Weigh 1g of waxy cornstarch, dissolve it in 100mL buffer solution, stir evenly, gelatinize in a boiling water bath, add a certain amount of pullulanase to debranch and enzymatically hydrolyze the starch for 6 hours at 58°C, centrifuge to take the supernatant, add the supernatant The mass concentration of the solution was 1‰Tween 80 (w / v) and the starch enzymatic solution was placed at 4°C for 12 hours to regenerate starch nanoparticles to prepare starch nanoparticles. The obtained starch nanoparticles were washed several times with distilled water and freeze-dried.

[0054] ② Preparation of low activity α-amylase

[0055] Weigh 10 mg of starch nanoparticles and dissolve them in 50 mL of phosphate buffer solution (pH 6.0), and ultrasonically disperse them for 2 min; Starch nanoparticle / α-amylase composite system.

Embodiment 2

[0057] (2)①Preparation of retrograde starch nanoparticles

[0058] Weigh 1g of waxy cornstarch, dissolve it in 100mL buffer solution, stir evenly, gelatinize in a boiling water bath, add a certain amount of pullulanase to debranch and enzymatically hydrolyze the starch for 7 hours at 55°C, centrifuge to take the supernatant, add the supernatant The mass concentration of the solution was 0.5‰ Tween 80 (w / v) and the starch enzymatic solution was placed at 4°C for 11 hours to regenerate starch nanoparticles to prepare starch nanoparticles. The obtained starch nanoparticles were washed several times with distilled water and freeze-dried.

[0059] ② Preparation of low activity α-amylase

[0060] Weigh 100 mg of starch nanoparticles and dissolve them in 50 mL of phosphate buffer solution (pH 6.0), and ultrasonically disperse them for 3 min; Starch nanoparticle / α-amylase composite system.

Embodiment 3

[0062] (2)①Preparation of retrograde starch nanoparticles

[0063] Weigh 1g waxy cornstarch, dissolve it in 100mL buffer solution, stir evenly, gelatinize in a boiling water bath, add a certain amount of pullulanase to debranch and enzymatically hydrolyze the starch for 5 hours at 60°C, centrifuge to take the supernatant, add the supernatant The mass concentration of the solution was 0.8‰Tween 80 (w / v) and the starch enzymatic solution was placed at 4°C for 13 hours to regenerate starch nanoparticles to prepare starch nanoparticles. The obtained starch nanoparticles were washed several times with distilled water and freeze-dried.

[0064] ② Preparation of low activity α-amylase

[0065] Weigh 100 mg of starch nanoparticles and dissolve them in 50 mL of phosphate buffer solution (pH 6.0), and ultrasonically disperse them for 4 min; Starch nanoparticle / α-amylase composite system.

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Abstract

The invention discloses a method for inhibiting the activity of amylase. The method is characterized in that starch nano-particles are prepared through a retrogradation technology and an anti-solventtechnology, and are used as a material for inhibiting the activity of alpha-amylase, that is starch and chitin are used to prepare the nano-particles, and the alpha-amylase is adsorbed by the nano-particles at a most suitable pH value in order to inhibit the activity of the alpha-amylase. The highly-stable and uniformly-distributed starch nano-particles and chitin whiskers are adopted to adsorb the alpha-amylase, and the starch nano-particles are rapidly combined with the active sites of the alpha-amylase to form an alpha-amylase-starch nano-particle compound in order to obstruct the combination of the alpha-amylase and soluble starch, so the content of the aphla-amylase combined with the soluble starch substrate is reduced, thereby the activity of the alpha-amylase is inhibited. The method for preparing alpha-amylase having low activity has the advantages of simplicity, easiness in operation, low cost, and suitableness for large-scale production, and is applied to the food and medicine industries.

Description

technical field [0001] The invention belongs to the technical field of improving enzyme activity of food and medicine, and in particular relates to a method for inhibiting the activity of amylase. Background technique [0002] With the continuous improvement of people's living standards, more and more people are troubled by diabetes, hypertension, obesity, etc., and people's health is seriously threatened. Among them, the prevalence of type 2 diabetes is getting higher and higher, and tends to be younger. Diabetes is becoming one of the important social problems affecting people's health and quality of life. Type 2 diabetes is a systemic disease mainly caused by elevated blood sugar due to insufficient insulin secretion in the body. According to statistics from the Ministry of Health, the prevalence rate of type 2 diabetes among adults over the age of 20 in my country is as high as 9.7%, and the prevalence rate in the early stage is as high as 15.5%. According to the lates...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/24C12N9/98C08B30/12C08B37/08
CPCC08B30/12C08B37/003C12N9/2414C12N9/99C12Y302/01001
Inventor 孙庆杰熊柳姜岁岁卢浩李曼赵梅
Owner QINGDAO AGRI UNIV
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