Production technology for fermentatively producing proteinase K by utilizing fungi microorganisms

A technology of microbial fermentation and production process, applied in the field of microbial fermentation of proteinase K, can solve the problems of low unit output, application limitation, high production cost, etc., and achieve the effect of meeting market demand

Inactive Publication Date: 2013-06-26
上海林叶生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Although proteinase K has a wide range of uses, its application is greatly restricted due to the low unit yield of microbial fermentation production, resulting in high production costs.

Method used

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  • Production technology for fermentatively producing proteinase K by utilizing fungi microorganisms

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Effect test

Embodiment 1

[0055] adopted through 60 The mutant strain IMU-112 obtained by Go gamma ray mutagenesis and protoplast ultraviolet mutagenesis is the starting bacterium. Pick 2 rings of the bacterium preserved on the slant with an inoculation loop and put it into a 1000ml Erlenmeyer flask containing 200ml of primary seed medium , A total of 3 bottles were inoculated. Regulate medium pH to be 5.5, shaker speed 180rev / min, after cultivating 44h at 28 ℃, inoculate in the 10L fermentor that 6 liters of secondary seed mediums are housed with 8% inoculum size, in the cultivation process, Control pH to 5.5, temperature to 28°C, ventilation rate to 1.2vvm, stirring speed to 350rev / min, continuous culture for 44 hours, then inoculate 7.5% inoculum into a 100L fermenter with a liquid capacity of 80L, and control the fermentation pH The temperature was 5.5, the temperature was 28°C, the ventilation rate was 1.0vvm, and the stirring speed was 350rev / min, and the fermented liquid was harvested for 64 ho...

Embodiment 2

[0057] adopted through 60 The mutant strain IMU-112 obtained by Go gamma ray mutagenesis and protoplast ultraviolet mutagenesis was selected as the starting bacterium. Pick 2 rings of the bacterium preserved on the slant with an inoculation loop and put it into a 1000ml Erlenmeyer flask containing 150ml of primary seed medium. , A total of 4 bottles were inoculated. Regulate medium pH to be 5.8, shaker speed 190rev / min, after cultivating 40h at 26 ℃, inoculate in the 10L fermentor that 5 liters of secondary seed culture medium are housed with 10% inoculum size, in the cultivation process, Control pH to 5.8, temperature to 26°C, ventilation rate to 1.2vvm, stirring speed to 280rev / min, continuous culture for 40h, and then inoculate 6% of the inoculum into a 100L fermenter with a liquid volume of 85L, and control the fermentation pH The temperature was 5.8, the temperature was 26°C, the ventilation rate was 1.0vvm, and the stirring speed was 400rev / min, and the fermentation liq...

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Abstract

The invention relates to a production technology for fermentatively producing proteinase K by utilizing fungi microorganisms. The production technology comprises the following steps of: using tritirachium album limber (Tritirachium Album Limber) as an original strain; mutagenizing by means of 60Go Gamma rays, and also mutagenizing by a protoplast in the presence of ultraviolet rays; introducing a special culture medium; and carrying out submerged fermentation to produce the proteinase K. Under the culture conditions that the culture temperature is 22 to 28 DEG C, the pH (Potential Of Hydrogen) of fermentation liquor is maintained within the range of 5.5 to 6.0, and a fermentation tank is at the speed of stirring of 250 to 500rev / min, the output of the proteinase K is greatly increased; and the production technology is suitable for being applied to industrial production. Compared with the prior art, the production technology for fermentatively producing the proteinase K by utilizing the fungi microorganisms, provided by the invention, can be used for filling the gap in production technology of the proteinase K of domestic, greatly increasing the biological expression quantity, and decreasing the production cost at the same time, and therefore, the market competitiveness of products can be greatly improved.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation of proteinase K, in particular to a production process for producing proteinase K by fermentation of fungal microorganisms. Background technique [0002] Proteinase K is a serine protease with broad cleavage activity, which is purified from the fermentation broth of Candida albicans (Tritirachium album limber). It cleaves the carboxy-terminal peptide bonds of aliphatic and aromatic amino acids for general degradation of proteins in biological samples. Because proteinase K is stable in urea and SDS and has the ability to degrade native proteins, it has a wide range of applications, including the preparation of chromosomal DNA for pulse electrophoresis, western blotting, and the removal of nucleases from DNA and RNA preparations. [0003] Although proteinase K has a wide range of uses, its application is greatly restricted due to the low unit yield of microbial fermentation productio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/58C12R1/645
Inventor 林克叶昀杨晓辉王洪森
Owner 上海林叶生物科技有限公司
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