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Variable-temperature production method for high-temperature-resistant fungi alpha-amylase

A technology of high temperature resistance and amylase, which is applied in the field of enzyme preparation, can solve the problems of unsuitable reaction process, low thermal stability, application and development limitations, etc., and is suitable for industrial production, strong stability, good stability and preservation active effect

Inactive Publication Date: 2016-04-27
湖南新鸿鹰生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the optimum action temperature of fungal α-amylase is around 55°C. When the temperature is higher than 60°C, it will gradually inactivate, and its thermal stability is relatively low, so it is not suitable for the reaction process at higher temperatures. Therefore, its application and development subject to certain restrictions

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A method for producing thermostable fungus α-amylase by variable temperature regulation, comprising the steps of:

[0023] (1) Activation of bacteria

[0024] Inoculate the well-preserved PDA slant strain of Trichoderma reesei 901-18 on the slant medium, culture at 27°C for 24 hours to activate the strain, and activate it twice;

[0025] (2) Expanded cultivation of liquid seeds

[0026] ①First-level seed culture: put the spore suspension made by activation in step (1) into a 500 ml shake flask, and make the spore concentration reach 10 5 cells / mL, medium capacity 100ml, rotary shaker 120r / min, culture temperature 27°C, culture time 10h;

[0027] ②Cultivation of secondary seeds: Inoculate 10% of the primary seeds into 500 ml secondary seed shake flasks, and the cultivation conditions are the same as those of the primary seeds;

[0028] ③Third-level seed cultivation: Inoculate the second-level seeds with a 10% inoculum amount into a 5000 ml third-level seed shake flask...

Embodiment 2

[0045] A method for producing thermostable fungus α-amylase by variable temperature regulation, comprising the steps of:

[0046] (1) Activation of bacteria

[0047] Inoculate the well-preserved PDA slant strain of Trichoderma reesei 901-18 on the slant medium, culture at 30°C for 30 hours to activate the strain, and activate it 3 times;

[0048] (2) Expanded cultivation of liquid seeds

[0049] ①First-level seed culture: put the spore suspension made by activation in step (1) into a 500 ml shake flask, and make the spore concentration reach 10 5 cells / mL, medium capacity 100ml, rotary shaker 150r / min, culture temperature 30°C, culture time 10h;

[0050] ②Cultivation of secondary seeds: Inoculate 10% of the primary seeds into 500 ml secondary seed shake flasks, and the cultivation conditions are the same as those of the primary seeds;

[0051] ③Third-level seed cultivation: Inoculate the second-level seeds with 10% inoculum amount into a 5000 ml third-level seed shake flask...

Embodiment 3

[0068] A method for producing thermostable fungus α-amylase by variable temperature regulation, comprising the steps of:

[0069] (1) Activation of bacteria

[0070] Inoculate the well-preserved slant strain of Trichoderma reesei 901-18 on the PDA slant medium, culture at 28°C for 36 hours to activate the strain, and activate it 3 times;

[0071] (2) Expanded cultivation of liquid seeds

[0072] ①First-level seed culture: put the spore suspension made by activation in step (1) into a 500 ml shake flask, and make the spore concentration reach 10 5 cells / mL, medium loading 100ml, rotary shaker 180r / min, culture temperature 28°C, culture time 15h;

[0073] ②Cultivation of secondary seeds: Inoculate 10% of the primary seeds into 500 ml secondary seed shake flasks, and the cultivation conditions are the same as those of the primary seeds;

[0074] ③Third-level seed cultivation: Inoculate the second-level seeds with 10% inoculum amount into a 5000 ml third-level seed shaker flask...

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PUM

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Abstract

The invention belongs to the field of enzyme preparations and discloses a variable-temperature production method for high-temperature-resistant fungi alpha-amylase. By gradient cooling and gradient heating type fermentation, anti-stress performance of 901-18Trichoderma reesei 901-18 is remarkably improved, enzymatic productivity of mixed cultures is presented maximally, and the high-temperature-resistant fungi alpha-amylase produced according to the method have the advantages of high enzyme activity, extensive action conditions, high stability and suitableness for industrial production.

Description

technical field [0001] The invention belongs to the technical field of enzyme preparation, in particular to a preparation method of high temperature resistant fungal alpha-amylase. Background technique [0002] The full name of α-amylase is α-1,4-glucan hydrolase (EC3.2.1.1). When acting on starch, it can cut α-1,4-glucosidic bonds from the inside of the molecule to generate dextrin and reduce Sugar, because the C1 carbon atom of the terminal glucose residue of the product is in the α-configuration, so it is named α-amylase. [0003] There are three common α-amylases: bacterial α-amylase, fungal α-amylase and malt α-amylase. Bacterial α-amylase is produced by bacteria such as Bacillus subtilis, Bacillus licheniformis, etc. Bacterial α-amylase has strong thermal stability. [0004] Fungal α-amylase was first extracted from Aspergillus oryzae by Gao Rangji of Japan in 1896 and used as a digestive agent. Since the 1950s, the scope of application of fungal α-amylase has gradu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/30C12R1/885
Inventor 李洪兵李海清张锦杰朱永明胡永明李贵骏
Owner 湖南新鸿鹰生物工程有限公司
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