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Method for extracting alpha-amylase inhibitor from white kidney beans by use of microwave-compound enzyme coupling method

An amylase inhibitor and compound enzyme technology, which is applied in the field of alpha-amylase inhibitors, can solve the problems of low purity and activity of alpha-amylase inhibitors, long time consumption, etc., so as to shorten enzymatic hydrolysis time, improve purity, The effect of shortening time

Inactive Publication Date: 2014-03-19
GUANGDONG FOOD IND INST
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In the prior art, the methods for extracting α-amylase inhibitors from white kidney beans are mostly time-consuming, and the purity and activity of α-amylase inhibitors are not high Therefore, the development of a method for rapidly and safely extracting pure natural, pure, and highly active α-amylase inhibitors from white kidney beans can promote the development and utilization of domestic agricultural product resources, and at the same time promote the development of the domestic hypoglycemic and weight loss market. Has a good application prospect

Method used

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  • Method for extracting alpha-amylase inhibitor from white kidney beans by use of microwave-compound enzyme coupling method

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Effect test

Embodiment 1

[0027] The method for extracting α-amylase inhibitors from white kidney beans by utilizing the microwave-complex enzyme coupling method provided in this embodiment comprises the following steps:

[0028] crude extraction : Take 1000g of white kidney beans and grind them at low temperature, add water to adjust the pH to 4.5 according to the ratio of 1:15 (mass ratio, the same below), then add 0.6% of the total weight of white kidney beans with glucoamylase and cellulase (glucoamylase: cellulase =10:1, mass ratio, the same below) placed under 700W, 55°C microwave radiation for 30min, wait until the temperature drops to room temperature and adjust its pH value to 6.5, then add papain and lipase of 0.3% of the total weight of white kidney beans (Papain:lipase=2:1, mass ratio, the same below) Place it under 700W, 50°C microwave radiation for 20min, then inactivate the enzymatic solution at 70°C for 30min;

[0029] refined : Centrifuge the filtrate after inactivating the enzyme,...

Embodiment 2

[0034] crude extraction : Take 1000g of white kidney beans and grind them at low temperature, add water at a ratio of 1:20 to adjust the pH to 4.5, then add 0.6% glucoamylase and cellulase (glucoamylase: cellulase = 15:1) and place in a 700W, 55℃ React under microwave radiation for 20 minutes. After the temperature drops to room temperature, adjust the pH value to 6.5, then add 0.3% papain and lipase (papain: lipase = 2:1) and place it under 600W, 50℃ microwave radiation for 30 minutes. , and then inactivate the enzyme at 70°C for 30min;

[0035] refined : Centrifuge the filtrate after inactivating the enzyme, ultrafilter the supernatant with an ultrafiltration membrane, and use CM-cellulose ion column chromatography on the retentate on the membrane, buffer with 0.025mol / L (pH4.5) acetic acid-sodium acetate solution (containing 0-1.0 mol / L NaCl) with an elution flow rate of 1mL / min, gradient elution, to collect the active protein peak, and the active eluate is filtered thr...

Embodiment 3

[0038] crude extraction : Take 1000g of white kidney beans and grind them at low temperature, add water at a ratio of 1:15 to adjust the pH to 4.5, then add 1.0% glucoamylase and cellulase (glucoamylase: cellulase = 18:1) and place in a 700W, 50℃ React under microwave radiation for 30 minutes, wait until the temperature drops to room temperature and adjust the pH value to 6.5, then add 0.3% papain and lipase (papain: lipase = 4:1) and place it under 700W, 55℃ microwave radiation for 30 minutes , and then inactivate the enzyme at 70°C for 30min;

[0039] refined : Centrifuge the filtrate after deactivating the enzyme, ultrafilter the supernatant with an ultrafiltration membrane, and use 0.025mol / L (pH4.5) acetic acid-sodium acetate to filter the retentate on the membrane through Sephadex G-75 gel column chromatography The buffer solution (containing 0-0.3 mol / L NaCl) was eluted at an elution flow rate of 1 mL / min, gradient elution, and the active protein peak was collected....

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Abstract

The invention discloses a method for extracting an alpha-amylase inhibitor from white kidney beans by use of a microwave-compound enzyme coupling method. According to the method, the white kidney beans are used as raw materials; the alpha-amylase inhibitor in the white kidney beans can be dissolved and separated out from the white kidney beans in an acceleration manner through microwave-compound enzyme coupling treatment after the white kidney beans are smashed; meanwhile, impurities such as fat, proteins and saccharides can be removed from the white kidney beans by only dozens of minutes through enzyme hydrolysis by one step under microwave-compound enzyme coupling catalytic hydrolysis, so that the purity of the alpha-amylase inhibitor in a coarse extract is much higher than that of the alpha-amylase inhibitor extracted by the common coarse extraction method; a pure high-activity alpha-amylase inhibitor can be obtained through ultrafiltration and ion column chromatographic refining. According to the method disclosed by the invention, the extraction condition is mild; water is used as a solvent, so that the pure nature of the product can be guaranteed; furthermore, due to the microwave-compound enzyme coupling method, the time for extracting the pure product is greatly shortened, and the inhibition activity of the product is over 90 percent; basic raw materials can be supplied to promotion of the development of a sugar reduction and fat reduction market in China.

Description

technical field [0001] The invention belongs to the technical field of α-amylase inhibitors, in particular to a method for extracting α-amylase inhibitors from white kidney beans by using a microwave-composite enzyme coupling method. Background technique [0002] The incidence of obesity is increasing year by year with the continuous improvement of people's living standards and unreasonable diet structure. It is listed as the world's four major medical and social problems along with AIDS, drug addiction and alcoholism, and seriously threatens human health. How to take effective measures to scientifically prevent and treat obesity is a hot topic that is widely concerned both at home and abroad. Obesity not only affects posture and activities, but also easily causes hyperlipidemia, atherosclerosis, coronary heart disease, high blood pressure, diabetes, fatty liver, gout, cholecystitis, and low resistance. China's current annual sales of weight-loss health care products have r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/48A61P3/10A61P3/04A61P3/06
Inventor 寇秀颖袁娟卫娜张素娟骆海平
Owner GUANGDONG FOOD IND INST
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