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Preparation method of modified alpha-amylase

An amylase and modification technology, applied in the field of biological science enzyme engineering, can solve problems such as unsolved accessibility, and achieve the effects of improving physical and chemical properties, enhancing freezing resistance, and improving thermal stability

Inactive Publication Date: 2017-05-24
SHANDONG SHENZHOU XIANGYU TECH GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the use of α-amylases with enhanced specific activity can only marginally improve starch hydrolysis since it does not address the issue of accessibility

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] A method for preparing modified α-amylase, the specific steps are as follows:

[0025] (1) Add 10 parts of xanthan gum to the protease aqueous solution for hydration. After hydration, the gum is squeezed and transferred to a homogeneous reactor for reaction. The reaction time is 3h, the reaction temperature is 30°C, and the reaction is inactivated after the end of the reaction. , Drying treatment, and then sieving the xanthan gum through a 120-mesh sieve, and reserve the rubber powder passing through the 120-mesh sieve for use; the drying method is drying, the drying temperature is 50℃, and the drying time is 3h;

[0026] (2) Place the ginger crushed to a particle size of 1mm in a container, add 2 times the weight of water, control the temperature at 50°C for 2 hours, add the mixture of 2 times the weight of ethanol and methanol, and control the temperature to 30°C for 5 hours , Filtration; The filtrate is concentrated in vacuo and freeze-dried to obtain ginger extract;

[00...

Embodiment 2

[0033] A method for preparing modified α-amylase, the specific steps are as follows:

[0034] (1) Add 12 parts of xanthan gum to the protease aqueous solution for hydration. After the hydration, the gum is squeezed and transferred to a homogeneous reactor for reaction. The reaction time is 3.5h and the reaction temperature is 35°C. Live and dry, and then siev the xanthan gum through a 120-mesh sieve, and reserve the rubber powder that passes through the 120-mesh sieve for use; the drying method is drying, the drying temperature is 55°C, and the drying time is 3.5h;

[0035] (2) Put the ginger crushed to a particle size of 1.5mm in a container, add 2.5 times the weight of water, control the temperature at 52°C for 2.2h, add the mixture material 2.2 times the weight of a mixture of ethanol and methanol, and control the temperature to 32°C Keep for 5.2h and filter; the filtrate is concentrated in vacuo and freeze-dried to obtain ginger extract;

[0036] (3) Dissolve potassium permangan...

Embodiment 3

[0042] A method for preparing modified α-amylase, the specific steps are as follows:

[0043] (1) Add 15 parts of xanthan gum to the protease aqueous solution for hydration. After hydration, the gum is squeezed and transferred to a homogeneous reactor for reaction. The reaction time is 4h, the reaction temperature is 40°C, and the reaction is inactivated after the completion of the reaction. , Drying, and then sieving the xanthan gum through a 120-mesh sieve, and reserve the rubber powder passing through the 120-mesh sieve for use; the drying method is drying, the drying temperature is 60℃, and the drying time is 4h;

[0044] (2) Place the ginger crushed to a particle size of 2mm in a container, add 3 times the weight of water, control the temperature at 55°C for 2.5 hours, add 2.5 times the weight of the mixture of ethanol and methanol, and control the temperature to 35°C to maintain 5.5h, filter; the filtrate is vacuum concentrated and freeze-dried to obtain the ginger extract;

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Abstract

The invention discloses a preparation method of modified alpha-amylase. The preparation method comprises the following concrete steps of adding an xanthan gumin into a protease water solution for hydratization, extruding the gum and shifting into a homogeneous reactor for reacting after finishing hydratization, inactivating and drying after finishing reaction, and screening for standby use; extracting a ginger root extract; preparing an oxidized xanthan gum solution; adding the ginger root extract into the oxidized xanthan gum solution; adding an alpha-amylase water solution into the solution, and then adding a reducing agent for reacting to obtain the modified alpha-amylase. According to the preparation method of the modified alpha-amylase provided by the invention, freezing-tolerant and heat-resisting performances of alpha-amylase can be improved at the same time, the same enzyme activity is maintained, the expiration date of the alpha-amylase is prolonged, the heat stability and the physical and chemical property of the prepared alpha-amylase are remarkably improved, the optimum pH (Potential of Hydrogen) value of the modified enzyme is more acidic than a free enzyme, and the enzyme activity can be less influenced by pH within a certain range.

Description

Technical field [0001] The invention relates to the field of biological science enzyme engineering, in particular to a preparation method of modified alpha-amylase. Background technique [0002] Bioenzyme is a new type of biocatalyst developed with the advancement of biotechnology. Compared with traditional catalysts, it has the advantages of high efficiency, low energy consumption and environmental friendliness. Alpha-amylase is also called starch-1,4-dextrinase (α-1,4-glucan glucanohydrolase, EC3.2.1.1), which is widely distributed in animals, plants, and microorganisms. Alpha-amylase has been extensively studied. The main function of α-amylase is to randomly cut α-1,4 glycosidic bonds from the inside of starch molecules to generate dextrin and reducing sugars. Therefore, its characteristic is to cause a sharp drop in the viscosity of the substrate solution and the disappearance of the iodine reaction. The main product is maltose when decomposing amylose. At present, α-amyla...

Claims

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Application Information

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IPC IPC(8): C12N9/96C12N9/26
CPCC12N9/96C12N9/2414C12Y302/01001
Inventor 任思凯李超周忠伟袁美宏任思龙
Owner SHANDONG SHENZHOU XIANGYU TECH GRP