Culture medium and method for producing insulin glargine precursor by fermenting with culture medium

Abstract

The invention relates to a culture medium and a method for producing an insulin glargine precursor by fermenting with the culture medium. The invention is characterized in that 1 liter of the culture medium comprises 3-8g of citric acid, 0.01-0.1g of ferric sulfate, 2-8g of diammonium hydrogen phosphate, 2-4g of monopotassium phosphate, 1-4g of magnesium sulfate, 8-15g of glucose, 1-3g of glycerinum, 10-16g of yeast extracts, 0.05-0.2g of vitamin B1, 0.5-1mg of trace element ammonium molybdate, 0.1-1mg of copper sulfate, 1-4mg of boric acid, 0.2-0.6mg of potassium iodide, 1-4mg of manganese chloride, 1-5mg of zinc acetate and the balance of water. The method comprises the steps that a recombinant escherichia coli BL21(DE3) / hp1 strain is cultured by using a solid slant culture medium to obtain a monoclone; the monoclone is inoculated onto a liquid seed culture medium to be cultured; the obtained liquid seed is inoculated on the culture medium; the insulin glargine precursor is prepared by oscillation fermentation culture by a shake flask or fermentation culture in a ventilation stirring fermentation tank. The method is easy to operate and low in cost of raw materials; the protein content of the obtained insulin glargine precursor reaches 7g / l at least and 11g / l at most, and a new path is opened up for large-scale production of the insulin glargine.

Description

technical field [0001] The invention designs a biological culture medium, specifically a new culture medium for fermenting and producing insulin glargine precursor, and a method for fermenting and producing insulin glargine precursor by using the culture medium. Background technique [0002] Insulin glargine is a genetically recombinant long-acting human insulin analogue, which belongs to the third generation of high-end insulin products. Insulin drugs have always had broad prospects for development. The development of insulin technology has gone through the extraction of animal insulin from the early animal pancreas to the biosynthesis of human insulin and human insulin analogues. The third generation of recombinant insulin analogs, insulin glargine, is the most high-end product of insulin. At present, Sanofi-Aventis' first third-generation high-end insulin "Come" has a global sales of nearly 5 billion US dollars in 2010, and it is the 7th "blockbuster" among the top 200 ...

AI Technical Summary

Problems solved by technology

In the fermentation of Escherichia coli, high-density fermentation under the condition of high-efficiency expression of exogenous genes is of great significance for reducing the culture volume, shortening the production cycle, improving production efficiency, reducing production costs, and simplifying the product purification process. However, high-density fermentation During the process, bacteria aging, autolysis and inability to express foreign genes are prone to occur.