Aspergillus oryzae bacterial strain giving high yield of neutral protease and liquid fermentation method thereof

A technology of neutral protease and Aspergillus oryzae, which is applied in the field of bioengineering, can solve the problems of viscous fermentation material liquid, affecting fermentation activity, etc., and achieve the effect of good heat resistance

Active Publication Date: 2013-04-03
SHANDONG LONGKETE ENZYME PREPARATION
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Problems solved by technology

Aspergillus oryzae liquid fermentation has the advantages of rich enzyme system, easy control of fermentation, and simple extraction process after fermentation, so it has received more and more attention. Liquid fer

Method used

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  • Aspergillus oryzae bacterial strain giving high yield of neutral protease and liquid fermentation method thereof
  • Aspergillus oryzae bacterial strain giving high yield of neutral protease and liquid fermentation method thereof
  • Aspergillus oryzae bacterial strain giving high yield of neutral protease and liquid fermentation method thereof

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Embodiment Construction

[0017] The present invention will be described in more detail below through specific examples, and the specific implementation cases are only for illustration, and are not intended to limit the implementation scope of the present invention.

[0018] Breeding of strains:

[0019] 1 Preparation of protoplasts

[0020] Take the fresh slopes of the two starting strains, wash the bacteria with sterile water, oscillate in a test tube with glass beads to disperse the bacteria, collect the bacteria by centrifugation, and use 0.6M sorbitol osmotic pressure stabilizer (0.1M citric acid buffer solution + 0.6M sorbitol + 0.01M EDTA) to suspend, centrifuge and wash, collect and suspend with osmotic pressure stabilizer, and store in ice for later use. Then add lysozyme 1.2%, helicase 0.8%, cellulase 0.8%, carry out enzymolysis at 32 ℃ for 6 hours, make protoplast standby, wherein the calculation method of protoplast preparation rate and regeneration rate is as follows:

[0021] Cultivate ...

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Abstract

The invention relates to an aspergillus oryzae mutant bacterial strain giving high yield of neutral protease and a liquid fermentation method thereof. The enzymatic activity of the neutral protease produced by the aspergillus oryzae bacterial strain is more than 20,821 u/mL. The liquid fermentation method for the neutral protease by using the aspergillus oryzae mutant bacterial strain CGMCC 6848 comprises the following steps: 1), seed culture; 2), liquid fermentation; and 3), extraction and refining of the neutral protease. According to the liquid fermentation method for the neutral protease, the fermentation activity is 25,000 u/mL, the residual enzyme activity is 91% after the produced neutral protease is subjected to heat preservation at the temperature of 40 DEG C for 1h, and the aspergillus oryzae mutant bacterial strain has good heat resistance and can be applied widely in industries such as the food industry, the feed industry and the textile industry.

Description

technical field [0001] The invention relates to an aspergillus oryzae mutant strain with high production of neutral protease and liquid fermentation technology thereof, belonging to the technical field of bioengineering. Background technique [0002] Neutral protease has mild action conditions. It is the earliest protease preparation discovered and widely used in industrial production. It can be widely used in food, feed, pharmaceutical and other industries. Now most of the neutral proteases in the market are produced by bacterial fermentation, and no one has been found. Method for producing neutral protease by liquid fermentation of Aspergillus oryzae. Aspergillus oryzae liquid fermentation has the advantages of rich enzyme system, easy control of fermentation, and simple extraction process after fermentation, so it has received more and more attention. Liquid fermentation of Aspergillus oryzae is affected by its phenotype. Unfavorable phenomena such as mycelium agglomerat...

Claims

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Application Information

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IPC IPC(8): C12N1/15C12N9/62C12R1/69
Inventor 刘顺启王兴吉李方方刘文龙
Owner SHANDONG LONGKETE ENZYME PREPARATION
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