Aspergillus oryzae strain and method for producing fungal alpha-amylase by liquid-state fermentation of aspergillus oryzae strain

A technology of liquid fermentation and Aspergillus oryzae, which is applied in the field of microbial fermentation, can solve the problems of expensive industrial production cost of fungal α-amylase, and achieve the effects of high yield, fast growth and simple process

Active Publication Date: 2014-09-10
意葆(上海)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the bottleneck of domestic fungal α-amylase production is the acquisition of high-yield strains and expensive industrial production costs

Method used

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  • Aspergillus oryzae strain and method for producing fungal alpha-amylase by liquid-state fermentation of aspergillus oryzae strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The mutagenesis screening of embodiment 1 Aspergillus oryzae bacterial strain of the present invention

[0035] The mutagenesis screening steps are as follows:

[0036] (1) The processing of single spore suspension: get the fresh slant (flat plate) that is coated with original bacterial strain (aspergillus oryzae strain that preservation number is CGMCC 1.1923) and add 10ml sterile saline, gently inoculate the spores on the slant with an inoculation needle Scrape off, pour into a sterile small Erlenmeyer flask with glass beads, oscillate to disperse the spores, filter with absorbent cotton; count with a microscope, and dilute the spore concentration to n×10 6 a / ml;

[0037] (2) Selection of mutagen dose: 5 mL of dispersed spore suspension was drawn into a sterile plate, placed in an ultraviolet mutagenesis box, and time doses of 5 s, 10 s, 15 s, 20 s, 25 s, and 30 s were respectively set for mutagenesis. After the mutagenesis is completed, spread after multiple diluti...

Embodiment 2

[0041] Embodiment 2 method of the present invention produces fungal α-amylase (50 liters of fermentation tanks)

[0042] 1. Culture medium [by weight and volume percentage (g / ml)]:

[0043] 1. Seed medium: 6% potato starch, 2% yeast powder, 0.3% dipotassium hydrogen phosphate, 0.3% magnesium sulfate heptahydrate, the balance is water, and the pH is 6.5;

[0044] 2. Fermentation medium in a 50-liter tank: 6% potato starch, 2% yeast powder, 0.3% dipotassium hydrogen phosphate, 0.3% magnesium sulfate heptahydrate, 0.01% soybean oil, the balance is water, and the pH is 6.5.

[0045] 2. Pretreatment

[0046] Strain culture:

[0047] Take an appropriate amount of Aspergillus oryzae strains with the preservation number CGMCC No.6834, and put them into four 500mL Erlenmeyer flasks, each containing 100ml of seed medium; culture conditions: 30°C, 200rpm shaker, culture for 48 hours, until the bacteria The liquid is viscous; use a portable all-stainless steel pulverizer to disperse th...

Embodiment 3

[0056] Embodiment 3 method of the present invention produces fungal α-amylase (10 tons of fermentation tanks)

[0057] 1. Culture medium [by weight and volume percentage (g / ml)]:

[0058] 1. Seed medium: 6% potato starch, 2% yeast powder, 0.3% dipotassium hydrogen phosphate, 0.3% magnesium sulfate heptahydrate, the balance is water, and the pH is 6.5;

[0059] 2. Fermentation medium: 4% potato starch, 6% soybean flour, 1% ammonium sulfate, 0.3% dipotassium hydrogen phosphate, 0.3% magnesium sulfate heptahydrate, 0.03% soybean oil, the balance is water, pH 6.5

[0060] 2. Pretreatment

[0061] 1. Strain cultivation:

[0062] Take an appropriate amount of Aspergillus oryzae strains with a preservation number of CGMCC No.6834, and insert them into five 1000mL Erlenmeyer flasks, each of which contains 200ml of seed medium; culture conditions: 30°C, 200rpm shaker culture, culture for 48 hours, Until the bacterial liquid is thick; use a portable all-stainless steel pulverizer to ...

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Abstract

The invention discloses an aspergillus oryzae strain MFA-AO-3 with the collection number of CGMCC No. 6834 and a method for producing fungal alpha-amylase by taking the aspergillus oryzae strain as the strain for liquid-state fermentation. The aspergillus oryzae strain can be used for realizing high yield of the fungal alpha-amylase; and the method for producing the fungal alpha-amylase by liquid-state fermentation of the aspergillus oryzae strain, disclosed by the invention, comprises the following step: on the basis of taking the aspergillus oryzae strain as the strain, performing fermentation and culture at the stirring rate of 500-1000 rpm (revolutions per minute) under the conditions that the ventilation quantity calculated by feed liquid volume is as follows: the air volume per minute is 1:(1.5-2) and the dissolved oxygen accounts for 20-25%, thereby preparing the fungal alpha-amylase. According to the method for producing the fungal alpha-amylase, disclosed by the invention, the enzyme production level is high and the industrial benefits are great.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation, in particular to an aspergillus oryzae strain and a method for producing fungal alpha-amylase by liquid fermentation thereof. Background technique [0002] α-amylase is an important enzyme preparation. It acts on the interior of starch to cut α-1,4 glycosidic bonds to generate dextrin and reducing sugar. The carbon atom configuration of the terminal residue of the product is α configuration, so Called α-amylase. The α-amylase in flour has obvious influence on the quality of fermented flour products. The content of α-amylase in flour processed from wheat harvested in normal seasons is generally insufficient. In order to improve and improve the quality of flour-based foods, α-amylase can be added during flour processing. [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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N9/30C12R1/69
Inventor 姜黎黎张淼李平顺
Owner 意葆(上海)生物科技有限公司
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