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Method for efficiently extracting mung bean alpha-amylase inhibitor

An amylase inhibitor and mung bean technology, which is applied in the extraction field of mung bean alpha-amylase inhibitor, can solve the problems of centrifugal separation operation obstacle, reduce the biological activity of the product, increase the viscosity of the solution, etc. Development prospects, easy operation and control effects

Active Publication Date: 2013-12-11
保定鲜尔康生物工程有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many domestic studies on extracting α-AI from wheat, but there is no report on extracting α-AI from mung bean, especially those with high inhibitory activity.
[0005] In addition, in the prior art, α-AI is mostly prepared by the traditional heating solvent extraction method. During the extraction process, it is heated to above 53°C, which will easily lead to gelatinization of part of the starch and increase the viscosity of the solution. obstacles; and prolonged heating can reduce the biological activity of the product
During the extraction process, due to process limitations, the raw materials cannot be crushed too finely, resulting in a decrease in the degree of fragmentation of the cell wall, which is not conducive to the complete release of α-AI in the cells, and requires a large amount of extraction solvent for long-term extraction.
Therefore, the traditional heating solvent extraction method has the disadvantages of long extraction time and high production cost when preparing α-AI.

Method used

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  • Method for efficiently extracting mung bean alpha-amylase inhibitor
  • Method for efficiently extracting mung bean alpha-amylase inhibitor
  • Method for efficiently extracting mung bean alpha-amylase inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Soak 5×0.1kg of defatted mung bean flour at 40°C for 4h, 8h, 12h, 16h, and 20h respectively, and then use a high-speed universal grinder to grind them into powders below 100 mesh. After stirring, centrifuge at 12,000g at 4°C for 30min to remove slag ;

[0040] Gel column SephedexG-75 extraction: Use 0.02mol / L, pH 4.0 HAc-NaAc buffer to equilibrate the gel column, then add the supernatant obtained after the previous step of centrifugation into HAc-NaAc buffer to a final concentration of 0.02 mol / L, load the sample at a flow rate of 0.5mL / min, then elute and extract with 0.02mol / L, pH 4.0 HAc-NaAc buffer, collect protein peaks and measure amylase inhibitory activity tube by tube, collect activity peaks and elute solution, mixed with ethanol pre-placed at -20°C according to the volume ratio of 1:1, and precipitated at -20°C. The extraction time is the time for the first Sephedex G-75 extraction to collect the active peak eluate and ethanol precipitation at -20°C. , is 1.5...

Embodiment 2

[0045] Soak 5×0.1kg of defatted mung bean powder in 30°C, 35°C, 40°C, 45°C, 50°C for 12 hours, and then use a high-speed universal grinder to make a powder with a mesh size of less than 100. After stirring, 12000g, 4°C , centrifuged for 30min to remove slag;

[0046] Gel column SephedexG-75 extraction: Use 0.02mol / L, pH 4.0 HAc-NaAc buffer to equilibrate the gel column, then divide the supernatant obtained after the previous step of centrifugation into five equal parts, add HAc-NaAc buffer to each solution to a final concentration of 0.02mol / L, loaded at a flow rate of 0.5mL / min, and then eluted with 0.02mol / L, HAc-NaAc buffer solution with a pH of 4.0 to collect protein peaks and measure amylase inhibition tube by tube. The active peak eluate was collected, mixed with ethanol pre-placed at -20°C in a volume ratio of 1:1, precipitated at -20°C, and the extraction time was the first Sephedex G-75 extraction to collect the active peak eluate and The time for ethanol precipitati...

Embodiment 3

[0051] Soak 5×0.1kg of defatted mung bean flour at 40°C for 12 hours, then use a high-speed universal grinder to make a mill powder with a mesh size below 100 mesh, and centrifuge at 12,000g at 4°C for 30 minutes to remove slag after stirring;

[0052] Gel column SephedexG-75 extraction: Use 0.02mol / L, pH 4.0 HAc-NaAc buffer to equilibrate the gel column, then divide the supernatant obtained after the previous step of centrifugation into five equal parts, add HAc-NaAc buffer to each solution to a final concentration of 0.02mol / L, loaded at a flow rate of 0.5mL / min, and then eluted with 0.02mol / L, HAc-NaAc buffer solution with a pH of 4.0 to collect protein peaks and measure amylase inhibition tube by tube. The active peak eluate was collected, mixed with ethanol pre-placed at -20°C in a volume ratio of 1:1, precipitated at -20°C, and the extraction time was the first Sephedex G-75 extraction to collect the active peak eluate and The time for ethanol precipitation at -20°C is 0...

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Abstract

The invention discloses a method for efficiently extracting a mung bean alpha-amylase inhibitor. According to the method, the alpha-amylase inhibitor with uniform ingredients is extracted and purified from mung bean by adopting gel column chromatography and ethanol precipitation, meanwhile, the influence of mung bean alpha-amylase inhibitor extraction conditions on the inhibitory activity of Porcine pancreatic alpha-amylase is analyzed, and the best extraction process conditions of the mung bean alpha-amylase inhibitor are determined. The mung bean alpha-amylase inhibitor extracted according to the method disclosed by the invention has high temperature stability and accordingly has a broad application prospect in the development of safe and natural blood glucose reducing drugs.

Description

technical field [0001] The invention relates to a method for extracting mung bean alpha-amylase inhibitor, in particular to a method for extracting mung bean alpha-amylase inhibitor known for its high efficiency and high product temperature stability. Background technique [0002] With the improvement of people's living standards and the prolongation of life expectancy, the number of patients with diseases such as obesity and diabetes is increasing year by year in both developed and developing countries, and related research on weight loss and hypoglycemia has received more and more attention. Since mung bean α-amylase inhibitor (α-Amylase Inhibitor, α-AI) is a glycoside hydrolase inhibitor, it hinders the digestion and absorption of carbohydrates in food by inhibiting the activity of saliva and pancreatic amylase in the intestinal tract. Sexually reduce sugar intake, lower blood sugar and blood lipid content, reduce fat synthesis, and then play a role in lowering blood suga...

Claims

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Application Information

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IPC IPC(8): A61K36/48A61P3/10
Inventor 王琦王荣侯沁文雷海英
Owner 保定鲜尔康生物工程有限责任公司
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