Method for increasing fermentation yield of plasmids

Abstract

The invention discloses a method for improving the fermentation yield of plasmids, which comprises the following steps: after fermenting for a certain time by using a primary fermentation culture medium, carrying out exponential fed-batch fermentation at a limited specific growth rate by using a fed-batch culture medium A so as to improve the yield of escherichia coli plasmids. According to the method for increasing the fermentation yield of the plasmids, by optimizing the material supplementing culture medium and the material supplementing method, high-quality and high-yield plasmid DNA can be obtained more quickly and more conveniently, high-density culture of escherichia coli is achieved, the plasmid yield is increased, and meanwhile the integrity of the plasmids can be kept.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation, in particular to a method for increasing the yield of plasmid fermentation. Background technique [0002] Plasmid DNA plays a key role in genetic engineering, genetically modified organisms, gene therapy, agriculture, and DNA vaccines. In recent years, with the continuous development of biological products, plasmid DNA is being increasingly used in new gene therapy, including DNA vaccines, gene replacement vectors, and viral vector production. The ability to consistently produce large quantities of high-purity, high-quality plasmid DNA has also become increasingly important. At present, plasmid DNA is mainly fermented by Escherichia coli, and obtained through a series of downstream processes such as extraction and chromatography (such as anion exchange and hydrophobic interaction). Among them, strain fermentation plays a dominant role in the production of plasmid DNA. In the pri...

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Problems solved by technology

In the prior art, shake flasks can only be used for the production of milligram-level plasmids, which cannot guarantee the quantity and quality of plasmids used in preclinical and clinical research, resulting in the inability to realize industrial scale production

[0003] At present, using Escherichia coli with high-yielding plasmids, using methods such as low-temperature culture and maintaining low growth rates, the bacteria are harvested after long-term cultivation, and the plasmids are extracted and purified. Although the yield is high, the fermentation cycle is too long, and the host protein Some host bacteria accumulate too much of their own products, which will affect the quality of the final product plasmid DNA

[0004] To sum up, in the existing plasmid DNA process production, the quality of the plasmid DNA product is still not good enough, and the process method cannot meet the requirements of economy and scale

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