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Fungal alpha-amylase and preparation method thereof

A technology of amylase and fungus, applied in the field of fungal α-amylase and its preparation, can solve the problems of pollution, low thermal stability, unsuitable treatment process, etc., achieve convenient process operation, stable activity, and simplify starch hydrolysis process operation Effect

Inactive Publication Date: 2009-06-10
SHANDONG LONGKETE ENZYME PREPARATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its thermal stability is low and it is not suitable for higher temperature processing. For example, if the brewing of industrial syrup is carried out at about 60°C, the amount of enzyme added must be doubled, and pollution problems may occur at 55°C

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] a. Pre-cultivation of liquid strains: Put 60 kg of glucose, 3 kg of peptone, 6 kg of corn steep liquor, 3.6 kg of bean cake powder, 0.15 kg of calcium chloride, and 0.99 kg of ammonium sulfate into a 1000L reactor, and add water to a total amount of 300 kg. , and adjust the pH to 6.0; raise the temperature to 121°C, and insulate and sterilize for 35 minutes. Cool to 30°C, insert Aspergillus oryzae; 30°C-40°C, ventilation volume 45m 3 / h, the pressure in the tank is 0.05Mpa-0.08Mpa, and the culture period is 32 hours to obtain liquid strains for solid fermentation;

[0030] b. Solid fermentation culture: 3000 kg of wheat bran, steamed for 60 minutes at a temperature of 121°C; cooled to 35°C, added the liquid strain prepared in step a diluted with water under stirring, to make the initial water content of the fermentation raw material At 40%-45%; at a temperature of 35°C, the ventilation rate is 60m 3 / h, fermented for 5.0 days, fermentation level: 14000u / g (dry koji). ...

Embodiment 2

[0033] a. Pre-cultivation of liquid strains: put 30 kg of starch, 3 kg of peptone, 6 kg of corn steep liquor, 3.6 kg of bean cake powder, 0.15 kg of calcium chloride, and 0.99 kg of ammonium chloride into a 1000L reactor, and add water to a total amount of 300 kg, and adjust the pH to 6.0; raise the temperature to 121°C, and keep the temperature for sterilization for 35 minutes. Cooled to 30°C, inserted into Aspergillus oryzae and passed the primary screening on the plate, and then used one of ultraviolet rays, ion beams, cobalt 60γ-rays, diethyl phosphate mutagens or any two or more of them to alternately mutagenize the strains ;30°C—40°C, ventilation volume 60m 3 / h, the pressure in the tank is 0.05Mpa-0.08Mpa, and the culture period is 32 hours to obtain liquid strains for solid fermentation;

[0034] b. Solid fermentation culture: 3000 kg of wheat bran, steamed for 50 minutes at a temperature of 123°C; cooled to 35°C, added the liquid strain prepared in step a diluted wit...

Embodiment 3

[0037] a. Pre-cultivation of liquid bacteria: 30 kg of maltose syrup, 6 kg of corn steep liquor, 3.6 kg of bean cake powder, 0.15 kg of calcium chloride, and 0.99 kg of ammonium chloride are put into a 1000L reactor, and water is added to a total amount of 300 kg, and adjusted PH to 5.6; heat up to 121°C, keep warm and sterilize for 35 minutes. Cooled to 30°C, inserted into Aspergillus oryzae and passed the primary screening on the plate, and then used one of ultraviolet rays, ion beams, cobalt 60γ-rays, diethyl phosphate mutagens or any two or more of them to alternately mutagenize the strains ;30°C—40°C, ventilation volume 60m 3 / h, the pressure in the tank is 0.05Mpa-0.08Mpa, and the culture period is 28 hours to obtain liquid strains for solid fermentation;

[0038] b. Solid fermentation culture: 3000 kg of wheat bran, steamed for 50 minutes at a temperature of 123°C; cooled to 35°C, added the liquid strain prepared in step a diluted with water under stirring, so that the...

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Abstract

The invention relates to fungus alpha-amylase and a preparation method thereof. The fungus alpha-amylase belongs to alpha-amylase from microbial sources. The preparation method is characterized in that an Aspergillus oryzae strain is subjected to liquid strain pre-culture and solid fermentation main culture; and the fungus alpha-amylase is prepared by immersion extraction, ultrafiltration concentration, dilution purification, ultrafiltration and vacuum drying, wherein the optimum pH of the fungus alpha-amylase is between 4.0 and 6.0, the proper temperature is between 45 and 65 DEG C, and the enzyme activity reaches more than 16,000 u / g (as calculated by dry medium). The invention provides the fungus alpha-amylase and the preparation method thereof, wherein the fungus alpha-amylase has high activity, high purity and higher thermal stability under the condition that the optimum pH is lower, is suitable for the prior amylolysis and saccharification technology, can simplify the amylolysis technological operation, stabilize the activity of the enzyme during production, improve the yield of sugar, and save water sources, is beneficial to environmental protection and is ideal. The fungus alpha-amylase is suitable for amylolysis and saccharification, and widely used for food fermentation and beverage processing, the fermentation industry, the enzyme preparation industry and the pharmaceutical industry.

Description

technical field [0001] The invention relates to a fungal alpha-amylase and a preparation method thereof. Belongs to alpha-amylases from microbial sources. Background technique [0002] α-amylase (α-1,4-glucan-4-glucoside hydrolase enzyme, EC3.2.1.1) from microbial sources is a class that can catalyze starch and other linear or branched 1.4-glucoside Enzyme for the hydrolysis of oligosaccharides and polysaccharides. It has high thermal stability and can catalyze the hydrolysis of α-1,4 bonds in macromolecules such as starch to produce maltodextrin with smaller molecular weight. Complete the liquefaction of starch segment bonds, so it is also called liquefying amylase. [0003] The traditional starch saccharification industry generally adopts double enzymatic method to hydrolyze starch to make sugar. That is, the starch is liquefied with α-amylase first, and then saccharified with glucoamylase to produce glucose. At present, the optimum pH of α-amylase on the market is 6....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/30C12R1/69
Inventor 王兴吉郭庆文王克芬
Owner SHANDONG LONGKETE ENZYME PREPARATION