Extraction method of insulin glargine precursor protein

Abstract

The invention relates to the field of biochemistry and discloses an extraction method of an insulin glargine precursor protein. The extraction method comprises the following steps of adding an acid into an insulin glargine precursor protein fermentation broth to adjust a pH value to 1-4, carrying out centrifugation and collecting a supernatant so that the insulin glargine precursor protein is obtained. Before bacterium centrifugation, the acid is used to adjust a pH value of the insulin glargine precursor protein fermentation broth to 1-4 so that after extraction, insulin glargine precursor protein content is improved, target protein loss caused by direct centrifugation removal of the bacteria is avoided and an insulin glargine yield is improved.

Description

[0001] This application claims the priority of a Chinese patent application filed with the Chinese Patent Office on November 21, 2012, the application number is 201210475070.3, and the invention title is "a method for extracting insulin glargine precursor protein", the entire content of which is incorporated by reference In this application. Technical field [0002] The invention relates to the field of biochemistry, in particular to a method for extracting insulin glargine precursor protein. Background technique [0003] In the treatment of diabetes, insulin has an irreplaceable role in controlling blood sugar standards and preventing complications. It is currently one of the most effective treatments for diabetes. In recent years, with the development of insulin technology, insulin analogues have been developed. One of the long-acting insulin analogues, insulin glargine, is being accepted and used by more and more doctors and patients. [0004] Insulin glargine is a biosynthetic...

AI Technical Summary

Problems solved by technology

Taking the adjustment of pH value as an example, existing reports such as US6218385 fermented broth add acid to inactivate the remaining microorganisms after fermentation, prevent broth autolysis and maintain a relatively low broth viscosity; CN200610011694. It is convenient to filter the capsule of bacteria; CN200710026682.3 is to add acid to the fermentation broth to prevent the target protein from being adsorbed by activated carbon; CN200810233835.6 is to add acid to the fermentation broth to prevent glutathione from being oxidized; Afterwards, the precipitate was diluted and acid was added to assist in dissolving natamycin, but none of the above reports involved the fermentation broth of insulin glargine precursor protein, and the starting point of adding acid was not to increase the protein content

At the same time, after the human insulin precursor fermentation broth was acidified before centrifugation, the recovery rate of its precursor protein was not significantly improved.

In addition, other reports of pretreatment of fermentation broth did not involve the increase of insulin glargine precursor protein content

[0007] As can be seen from this, there are almost no reports on how to improve the content of insulin glargine precursor protein in the extraction process in the prior art.

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