Method for extracting alpha-amylase inhibitor from hulless oat

A technology of amylase inhibitor and naked oats, which is applied in the field of extracting α-amylase inhibitor, can solve the problems of high price, side effects in clinical application, etc., and achieve the effect of high purity

Inactive Publication Date: 2010-11-24
SHANXI UNIV
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  • Description
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  • Application Information

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Problems solved by technology

The α-amylase enzyme inhibitor drugs currently on the market have a good effect of lowering postprandial blood sugar, but their price is high, and there ar

Method used

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  • Method for extracting alpha-amylase inhibitor from hulless oat
  • Method for extracting alpha-amylase inhibitor from hulless oat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: Preparation of α-amylase inhibitor

[0022] Crushing and degreasing:

[0023] Dehull naked oat seeds and crush them into naked oat flour; weigh 100g of naked oat flour, add 500ml of acetone, leave it at room temperature for 12 hours, and pump it into acetone powder under a vacuum decompression pump;

[0024] Protein extraction and rough separation: Dissolve acetone powder in 100ml of 20mM Tris-HCl pH8.0 buffer solution containing 100mM NaCl, then centrifuge the dissolved protein solution in a high-speed centrifuge at 12000rpm, 4°C for 30min, and take the supernatant Carry out ammonium sulfate precipitation, collect 30%-80% ammonium sulfate precipitate, suspend with 20ml of 20mM Tris-HCl pH8.0 buffer, put in a dialysis bag, dialyze with 1000ml of the same buffer, and replace the outer dialysate four times;

[0025] Protein column chromatography separation: The dialyzed protein solution was centrifuged in a high-speed centrifuge at 12000rpm at 4°C for 30min...

Embodiment 2

[0029] Example 2: Determination of oat α-amylase inhibitor activity and comparison with acarbose amylase inhibitory activity

[0030] 1. Determination of inhibitory activity of α-amylase inhibitors:

[0031] Use 3.5-dinitrosalicylic acid method for determination, incubate 100 μL α-amylase and 100 μL α-amylase inhibitor at 37°C for 10 min, then add 100 μL 1% soluble starch (containing 20 mM NaCl, 0.2mM CaCl 2 20mM sodium phosphate buffer pH 6.0), kept at 37°C for 10min, and finally added 1ml of 3.5-dinitrosalicylic acid to the reaction system, bathed in boiling water for 10min, made up the reaction system to 10ml with distilled water, and measured at 540nm Light absorption, without adding α-amylase inhibitor as a control.

[0032] The activity of an α-amylase inhibitor is defined as: the amount of the inhibitor required to inhibit α-amylase catalyzing the production of 1 μmol of glucose per minute at 37°C.

[0033] 2. Comparison of inhibitory activity of naked oat α-amylase ...

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Abstract

The invention discloses a method for extracting an alpha-amylase inhibitor from hulless oats. The method comprises the following steps of: decladding and pulverizing a hulless oat seed; degreasing with acetone; and then carrying out ammonium sulfate precipitation, ion-exchange chromatography and gel chromatography to obtain an electrophoretic pure alpha-amylase inhibitor, wherein the inhibiting specific activity is 26.43U/mg. A material for extracting the alpha-amylase inhibitor is derived from the hulless oat, a purifying method is simple and convenient, a product has high purity and high inhibiting activity, and the extracted alpha-amylase inhibitor can be used for preparing a medicament for reducing weight and treating diabetes mellitus.

Description

technical field [0001] The invention relates to a method for extracting plant effective components, in particular to a method for extracting alpha-amylase inhibitors from oatmeal seeds. Background technique [0002] In recent years, with the improvement of people's living standards, diabetes, high blood pressure, and obesity are becoming common high incidences, seriously endangering human health. Diabetes is a systemic disease mainly caused by the disorder of glucose metabolism caused by insufficient insulin secretion in the body. Obesity causes many diseases, such as hypertension, coronary heart disease, cardiovascular disease, diabetes, hyperlipidemia, etc., and becomes the biggest killer that threatens human health and life satisfaction. In clinical medicine, α-amylase inhibitors can prevent and treat diseases such as diabetes, hypertension, and hyperlipidemia. [0003] α-amylase inhibitors are one of the glycoside hydrolase inhibitors. They are a new class of oral anti...

Claims

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Application Information

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IPC IPC(8): C07K14/415C07K1/36C07K1/30C07K1/18C07K1/16
Inventor 石亚伟闫永飞钮利喜
Owner SHANXI UNIV
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