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Study of anthropogenic SOD expression by recombinant pichia pastoris

A Pichia pastoris, recombinant technology, applied in the biological field, can solve the problems of low SOD expression and low enzyme activity

Inactive Publication Date: 2016-08-17
SHENYANG MEDICAL COLLEGE
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, in the current research on the culture conditions of Pichia pastoris expression, after the fermentation process, there are unfavorable factors such as low SOD expression content, intracellular expression, and low enzyme activity.

Method used

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  • Study of anthropogenic SOD expression by recombinant pichia pastoris
  • Study of anthropogenic SOD expression by recombinant pichia pastoris
  • Study of anthropogenic SOD expression by recombinant pichia pastoris

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Experimental program
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Embodiment Construction

[0025] The technical scheme adopted in the present invention is:

[0026] 1. The optimized culture conditions in the study of Pichia pastoris recombinants expressing human SOD include:

[0027] 1) culture medium;

[0028] Recombinant Pichia pastoris was cultured in BMGY for induced expression;

[0029] 2) ventilation;

[0030] Cultivate BMGY with recombinant Pichia pastoris in a Erlenmeyer flask with a baffle on the bottom or side wall. The volume of the Erlenmeyer flask is in the range of 50-2000ml, and the volume of the medium must be less than or equal to 10% of the volume of the Erlenmeyer flask- 30%, use 2-3 layers of cotton gauze or a loose cork to cover the triangular bottle;

[0031] 3) temperature and shaking speed of the shaker;

[0032] Put the triangular flask containing the culture medium in a constant temperature shaking incubator and set the temperature at 30°C; if the constant temperature shaking incubator is a floor-standing constant temperature shaking in...

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Abstract

The invention discloses a study of of anthropogenic SOD expression by recombinant pichia pastoris. The enzyme activity of the recombinant pichia pastoris for anthropogenic SOD expression is the maximum and can reach 18.322 U / ml by optimizing the culture conditions, wherein the conical flask is 500 ml, the volume of liquid is 20%, the temperature is 30 DEG C, the oscillation revolving speed is 225 rpm, the Cu2+ concentration is 1%, the methanol concentration is 1%, and cultivation is performed by 24 h on the culture medium. After the fermentation conditions are optimized, secreting type SOD fermentation liquor can be obtained, and an SOD product with the enzyme activity can be obtained.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the research on the expression of human SOD by Pichia pastoris recombinants. Background technique [0002] As a eukaryotic cell, Pasteur4fswh8p has many advantages of expression system characteristic of higher eukaryotic cells, such as protein processing, protein folding, post-translational modification, and is as easy to operate as E. coli or yeast Saccharomyces cerevisiae organisms. Compared with baculovirus or mammalian tissue culture, Pichia pastoris has the advantages of faster, more convenient, cheaper, and high-level expression. As a type of yeast, Pichia pastoris has the same molecular and genetic manipulation advantages as Saccharomyces cerevisiae, but also has 10 to 100 times higher expression levels of heterologous proteins. These characteristics make it possible for Pichia pastoris to be used as a protein expression system. [0003] Pichia pastoris is able t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/81C12N9/02C12R1/84
CPCC12N15/81C12N9/0089C12N2800/102C12Y115/01001
Inventor 肖纯凌李新鸣毕延波张奕晶孙冶李舒音海晓欧
Owner SHENYANG MEDICAL COLLEGE
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