Biosynthesis of therapeutic DNA plasmid using agri-food subproducts media

• Autor(a) principal: Matos, Mafalda do Espírito Santo Silva
• Data de Publicação: 2015
• Tipo de documento: Dissertação
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10400.6/5937
• topic: Cheese Whey Powder Solution; Corn Steep Liquor; Dna Plasmídico; Escherichia Coli Vh35; Terapia Génica; Domínio/Área Científica::Engenharia e Tecnologia:: Outras Engenharias e Tecnologias

Increased relevance of biomedical applications of plasmid DNA (pDNA) to induce therapeutic effects has having a great impact on industry and biopharmaceutical research. Gene therapy and DNA vaccines are a new approach for treating genetic diseases such as cancer. These new therapies are based on delivery systems of drugs based on nucleic acids that act at the level of the target cell, correcting or deleting the affected cells. Escherichia coli (E. coli) is the most used host system in the production of pDNA, due to their high growth in minimal media and its easy genetic manipulation. In this paper we used E. coli VH35, a variant with genetic modifications that minimize the accumulation of acetate. Several studies have been performed in different steps of production and purification of plasmid DNA into E. coli in order to increase the yield and productivity of this biomolecule. However, despite several optimization achieved, commercial substrates are used as culture media to produce this biomolecule, taking into account that the final yield of pDNA make the whole production process quite expensive. In order to overcome this problem, the commercial substrates may be supplemented or replaced by industrial wastes resulting in a pDNA yield very similar compared to commercial substrates, via a more economical process. In this paper we used the CSL ("corn step liquor") and the CWPS ("Cheese whey power solution") as alternative substrates in culture media for production of pDNA in E. coli.