A method for preparing microplasmin without self-cutting form

A plasmin and microfiber technology is applied in the field of enzyme preparation to achieve the effects of reducing the use of material costs, improving market competitiveness and increasing yield

Active Publication Date: 2021-07-13
CHONGQING PEG BIO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] The present invention provides a method for preparing microplasmin without self-cutting, which solves the problem of not only avoiding the stability of microplasmin due to the acid hydrolysis factor of too low pH in the prior art, but also avoiding the problem caused by too high pH value. The problem of autocatalytic degradation of microplasmin

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  • A method for preparing microplasmin without self-cutting form
  • A method for preparing microplasmin without self-cutting form
  • A method for preparing microplasmin without self-cutting form

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Embodiment 2

[0065] Example 2: All the steps in the above-mentioned seed liquid culture and liquid fermentation culture were used to obtain a fermentation liquid (wherein, in step S1232, the concentration of (NH4)2SO4 was kept at 0.1 mol / L and sucrose was induced for 24 hours), and then the fermentation liquid was subjected to electrophoresis To detect the expression of the target protein, the volume ratio of the fermentation broth to the polyacrylamide gel was 85%:15% during electrophoresis.

Embodiment 3

[0066] Example 3: All the steps in the above-mentioned seed liquid culture and liquid fermentation culture were used to obtain a fermented liquid (wherein, in step S1232, the concentration of (NH4)2SO4 was kept at 0.25 mol / L and sucrose was induced for 24 hours), and then the fermented liquid was subjected to electrophoresis To detect the expression of the target protein, the volume ratio of the fermentation broth to the polyacrylamide gel was 85%:15% during electrophoresis.

Embodiment 4

[0067] Example 4: All the steps in the above-mentioned seed liquid culture and liquid fermentation culture were used to obtain a fermentation liquid (wherein, in step S1232, the concentration of (NH4)2SO4 was kept at 0.5 mol / L and sucrose was induced for 24 hours), and then the fermentation liquid was subjected to electrophoresis To detect the expression of the target protein, the volume ratio of the fermentation broth to the polyacrylamide gel was 85%:15% during electrophoresis.

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Abstract

The present invention proposes a method for preparing microfibrinolytic enzymes without self-cutting, including steps: S1, recombinant fermentation of human microfibrinolytic enzymes without self-cutting; step S1 includes steps: S12, liquid fermentation culture, in (NH4)2SO4 and sucrose were added during the liquid fermentation culture. The preparation method of microplasmin without self-cleavage solves the problem in the prior art that the acid hydrolysis of microplasmin due to too low pH can not be avoided while avoiding the self-catalytic degradation of microplasmin due to too high pH .

Description

technical field [0001] The invention relates to an enzyme preparation method, in particular to a method for preparing microplasmin without self-cleavage. Background technique [0002] Plasminolytic enzyme (plasmin) is a serine protease derived from plasminogen, an important component of mammalian blood. Human plasminogen (Plasminogen, Plg) is a multi-domain protein consisting of 791 amino acid residues, consisting of an N-terminal proactivation domain, 5 homologous Kringle domains (each about 80 amino acids), a The catalytic domain of serine protease and the connection sequence between domains, with a molecular weight of about 92,000 Daltons, is one of the key components of the human fibrinolytic system. Use plasmin to cut the N-terminal Arg68-Met69 of human plasminogen, or The peptide bond between Lys77-Lys78 or Lys78-Val79 produces a shortened zymogen called lysine-plasminogen. Further cleavage by elastase removes the first 4 Kringle domains to produce another zymogen, s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/68
CPCC12N9/6435C12Y304/21007
Inventor 杨辉何勇冷国政黄金凤潘武生范开
Owner CHONGQING PEG BIO BIOTECH CO LTD
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