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Preparation method of acid-resistant high-temperature-resistant alpha-amylase

A production method and high temperature resistant technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, enzymes, etc., can solve the problems of long fermentation cycle and achieve the effect of short cycle, high vitality and strong adaptability

Inactive Publication Date: 2016-08-17
温文伟
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are not many acid-resistant high-temperature α-amylases independently researched and produced in my country. The common OGO high-temperature-resistant acid α-amylase is refined from Bacillus licheniformis through liquid submerged fermentation and ultrafiltration. However, Its enzyme activity generally can only reach 20000 U / mL, and the fermentation period is long

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 The production method of the acidic high-temperature-resistant α-amylase is prepared by selecting genetically improved Bacillus subtilis through plate culture, shake flask culture, seed tank fermentation, fermentation tank fermentation, and enzyme extraction. It is characterized in that: the steps are as follows:

[0020] (1) Plate culture: medium formula: peptone: 1%, yeast powder: 0.2%, beef powder: 0.3%, sodium chloride: 0.2%, glucose: 5%, corn starch: 1%, agar powder: 1.7% , the balance is water, each component is expressed by weight percentage, the pH is controlled to be 6.7, and then sterilized at a temperature of 121°C and a pressure of 0.1Mpa for 30min, and incubated at a constant temperature of 36°C for 40h after inoculation;

[0021] (2) Shake flask culture: medium formula: 1.5% yeast powder, 0.5% glucose, 0.3% potassium dihydrogen phosphate, 0.1% magnesium sulfate, and the balance is water, and the components are expressed in weight percent; adjust ...

Embodiment 2

[0028] Example 2 The production method of the acidic high-temperature-resistant α-amylase is prepared by selecting genetically improved Bacillus subtilis through plate culture, shake flask culture, seed tank fermentation, fermentation tank fermentation, and enzyme extraction. It is characterized in that: the steps are as follows:

[0029] (1) Plate culture: medium formula: peptone: 0.5%, yeast powder: 0.1%, beef powder: 0.1%, sodium chloride: 0.1%, glucose: 3%, corn starch: 0.5%, agar powder: 1.5% , the balance is water, each component is expressed in weight percentage, the pH is controlled to be 6.5, and then sterilized at a temperature of 121°C and a pressure of 0.1Mpa for 30min, and incubated at a constant temperature of 36°C for 36h after inoculation;

[0030] (2) Shake flask culture: medium formula: 1% yeast powder, 0.3% glucose, 0.1% potassium dihydrogen phosphate, 0.05% magnesium sulfate, and the balance is water, and the components are expressed in weight percent; adj...

Embodiment 3

[0037] Example 3 The production method of the acidic high-temperature-resistant α-amylase is prepared by selecting genetically improved Bacillus subtilis through plate culture, shake flask culture, seed tank fermentation, fermentation tank fermentation, and enzyme extraction. It is characterized in that: the steps are as follows:

[0038] (1) Plate culture: medium formula: peptone: 1.5%, yeast powder: 0.3%, beef powder: 0.5%, sodium chloride: 0.3%, glucose: 7%, cornstarch: 1.5%, agar powder: 2% , the balance is water, each component is expressed in weight percentage, the pH is controlled to be 7.0, then sterilized at a temperature of 121°C and a pressure of 0.1Mpa for 30min, and incubated at a constant temperature of 36°C for 48h after inoculation;

[0039] (2) Shake flask culture: medium formula: 2% yeast powder, 0.7% glucose, 0.5% potassium dihydrogen phosphate, 0.15% magnesium sulfate, and the balance is water, and the components are expressed in weight percent; adjust the...

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Abstract

The invention discloses a preparation method of an acid-resistant high-temperature-resistant alpha-amylase. The preparation method includes selecting genetically-modified bacillus subtilis and subjecting the genetically-modified bacillus subtilis to plate culture, shake-flask culture, seed tank fermentation, fermentation tank fermentation and enzyme extraction so as to obtain the acid-resistant high-temperature-resistant alpha-amylase. The preparation method has the advantages that the genetically-modified bacillus subtilis cannot produce spores and is high in adaptability, less prone to contamination, high in activity and short in period; by adjusting air capacity control and culture medium ratio and effectively controlling a pH value in a preparation process, the prepared acid-resistant high-temperature-resistant alpha-amylase with the optimum reaction temperature of 90-100 DEG C, the pH value of 4.0-5.5, the enzyme activity of 40,000 U / mL and a fermentation period only lasting for 120 hours is a current domestic acid-resistant amylase maximum in enzyme activity within the shortest fermentation period.

Description

Technical field [0001] The present invention involves the production method of biological enzyme preparations, especially the production method of an acidic high temperature resistant alpha-amylase. Background technique [0002] α-amylase (α-1, 4-D-glucose-glucoside hydrolytase) is generally distributed in animals, plants and microorganisms, and is an important starch hydrolytic enzyme.When it acts on starch, the α-1 and 4 glycoside bonds are randomly cut from the internal of the starch molecule to generate paste and restore sugar.Because the end residual carbon atomic structure of the product is α configuration, it is called α-amylase.α-amylase is a very important enzyme preparation. It is used in a large amount of food processing, food industry, brewing, fermentation, textile industry and pharmaceutical industries. It is one of the most widely used enzyme preparations. [0003] The best response conditions of acid high temperature α-amylase generally are between 80-100 ° C and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/26C12R1/125
CPCC12N9/2414C12Y302/01001
Inventor 温文伟温明
Owner 温文伟
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