Optimization of culture medium for cell growth and expression of 648 antigen from Leishmania infantum chagasi in recombinant Escherichia coli M15

• Autor(a) principal: Santos, Everaldo Silvino dos
• Data de Publicação: 2014
• Outros Autores: Vaz, Michelle Rossana Ferreira, Sousa Junior, Francisco Canindé de, Costa, Letícia Maia Resende, Martins, Daniella Regina Arantes, Macedo, Gorete Ribeiro de
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UFRN
• DOI: 10.1007/s13213-014-1000-2
• Texto Completo: https://repositorio.ufrn.br/handle/123456789/32526
• topic: Culture medium; 648 antigen; Escherichia coli M15; Leishmania infantum chagasi

Visceral leishmaniasis is a potentially fatal disease caused by Leishmania infantum chagasi in the New World. Anti-leishmanial treatment is problematic because antileishmanial drugs are toxic and costly and drug resistance is emerging. Moreover, there is no effective and safe vaccine approved for human use against any form of leishmaniasis. It seems likely that both a subunit vaccine and an optimized diagnostic test would benefit from the production of recombinant antigens. However, for recombinant protein production, the composition of the cell growth medium must be carefully formulated and monitored, because it may have significant metabolic effects on both the cells and protein expression. Thus, the aim of this study was to optimize the culture medium for cell growth and the expression of the 648 antigen from Leishmania i. chagasi in recombinant Escherichia coli M15. The induction allowed the expression of the 648 antigen; this was confirmed by electrophoresis, which showed a protein with a molecular mass of 24 kDa. The results showed that the higher cell concentration obtained in TB medium can be explained by the larger number of components such as amino acids and vitamin precursors contained in this complex medium, as well as glycerol as a carbon and energy source. Acetic acid excretion, detected at the end of fermentation, resulted in an inhibitory effect for both cell growth and protein production, because this was above the concentration limit of 0.9 g L−1