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Method for industrially extracting alpha-amylase inhibitor in white kidney beans

The technology of an amylase inhibitor and white kidney bean is applied in the directions of pharmaceutical formulations, medical preparations containing active ingredients, metabolic diseases, etc., and can solve the problems of incapability of industrialized large-scale application, inability to obtain active alpha-amylase inhibitors, and the like, Achieve the effect of promoting comprehensive utilization, increasing added value, and helping prevention and control

Active Publication Date: 2015-04-22
无锡华康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The existing methods for extracting α-amylase inhibitors from white kidney beans rely on chromatographic columns, which cannot be applied on a large scale industrially, or cannot obtain highly active α-amylase inhibitors, and thus cannot meet the urgent needs of today's society

Method used

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  • Method for industrially extracting alpha-amylase inhibitor in white kidney beans
  • Method for industrially extracting alpha-amylase inhibitor in white kidney beans
  • Method for industrially extracting alpha-amylase inhibitor in white kidney beans

Examples

Experimental program
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Effect test

Embodiment 1

[0059] Take 100g white kidney beans and soak them in 300mL distilled water for 10h at room temperature, then manually peel them;

[0060] Use a high-speed tissue masher to grind the peeled white kidney beans with 300mL of distilled water and pass through a 100-mesh sieve, then use 300mL of distilled water to refine again and pass through a 100-mesh sieve. After standing for 2 hours, centrifuge at 9000r / min for 20 minutes at 4°C, and take the supernatant;

[0061] Inactivate the supernatant in a water bath at 70°C for 30 minutes, centrifuge at 9000 r / min for 20 minutes at 4°C, and collect the supernatant;

[0062] Add absolute ethanol to the supernatant to a final concentration of 70% (v / v), stir at 4°C for 2h, centrifuge at 9000r / min for 20min at 4°C, and collect the precipitate, which is protein;

[0063] Put the protein in the fume hood for 1 hour, and dissolve it with 300mL 0.02mol / L, pH6.9 phosphate buffer to obtain the protein solution;

[0064] After adjusting the conc...

Embodiment 2

[0070] Take 100g white kidney beans and soak them in 400mL distilled water for 8 hours at room temperature, then manually peel them;

[0071] Use a high-speed tissue masher to grind the peeled white kidney beans with 400mL of distilled water and pass through a 100-mesh sieve, then use 300mL of distilled water to refine again and pass through a 100-mesh sieve. After standing for 3 hours, centrifuge at 9000r / min for 20 minutes at 4°C, and take the supernatant;

[0072] Inactivate the supernatant in a water bath at 70°C for 30 minutes, centrifuge at 9000 r / min for 20 minutes at 4°C, and collect the supernatant;

[0073] Add absolute ethanol to the supernatant to a final concentration of 70% (v / v), stir at 4°C for 2h, centrifuge at 9000r / min for 20min at 4°C, and collect the precipitate, which is protein;

[0074] After the protein was placed in a fume hood for 2 hours, it was dissolved with 400 mL of 0.02 mol / L, pH 6.9 phosphate buffer to obtain a protein solution.

[0075] Aft...

Embodiment 3

[0081] Take 100g white kidney beans and soak them in 500mL distilled water for 7 hours at room temperature, then manually peel them;

[0082] Use a high-speed tissue masher to grind the peeled white kidney beans with 400mL of distilled water and pass through a 100-mesh sieve, then re-fine the sieve with 400mL of distilled water and pass through a 100-mesh sieve, discard the sieve, combine the filtrate, and statically After standing for 2 hours, centrifuge at 9000r / min for 20 minutes at 4°C, and take the supernatant;

[0083] Inactivate the supernatant in a water bath at 70°C for 30 minutes, centrifuge at 9000 r / min for 20 minutes at 4°C, and collect the supernatant;

[0084] Add absolute ethanol to the supernatant to a final concentration of 70% (v / v), stir at 4°C for 2h, centrifuge at 9000r / min for 20min at 4°C, and collect the precipitate, which is protein;

[0085] After the protein was placed in a fume hood for 1.5 h, it was dissolved with 200 mL of 0.02 mol / L, pH 6.9 pho...

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Abstract

The invention discloses a method for industrially extracting an alpha-amylase inhibitor in white kidney beans, belonging to the field of preparation of plant effective components. The method comprises the following steps: firstly adding water into soaked and peeled white kidney beans, grinding into slurry, sieving to remove residues, performing standing, centrifuging, enzyme deactivation, centrifuging and ethanol precipitation on an obtained filtrate to obtain protein, dissolving the obtained protein by using a phosphate buffer solution, performing moderate enzyme hydrolysis by using alkaline protease to change isoelectric points of impurity protein, then performing acid precipitation, collecting supernatant liquid, and performing ethanol precipitation and freeze drying to obtain an alpha-amylase inhibitor product. The prepared alpha-amylase inhibitor disclosed by the invention is high in purity and strong in activity; and moreover, the alpha-amylase inhibitor needs simple equipment, is low in cost, is pollution-free, has relatively good economic and environmental benefits, and can be applied to industrial large-scale production.

Description

technical field [0001] The invention relates to a method for industrially extracting alpha-amylase inhibitors from white kidney beans, which belongs to the field of preparation of plant active ingredients. Background technique [0002] With the development of social economy and the improvement of living standards, diabetes has gradually become a common disease, which poses a serious threat to human health. Diabetes is a systemic disease mainly characterized by disorders of glucose metabolism, and postprandial hyperglycemia is a typical feature of diabetes. The continuous rise of blood sugar after a meal will produce glucose toxicity, damage the cardiovascular system, and eventually cause irreversible damage to the human body. Therefore, controlling postprandial blood sugar rise is crucial for controlling and delaying the onset and development of diabetes. At present, there are more than 150 million diabetics in the world, and it is expected to reach 300 million by 2025. Am...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/48A61P3/10
CPCA61K36/48A61K2236/15A61K2236/19A61K2236/51A61K2236/53
Inventor 赵伟让一峰杨瑞金
Owner 无锡华康生物科技有限公司
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