| Texto completo | |
| Autor(es): |
Lefin, Nicolas
;
Miranda, Javiera
;
Munhoz Costa, Iris
;
Pedroso Reynaldo, Alejandro
;
Monteiro, Gisele
;
Zamorano, Mauricio
;
Pessoa Jr, Adalberto
;
Farias, Jorge G.
Número total de Autores: 8
|
| Tipo de documento: | Artigo Científico |
| Fonte: | FERMENTATION-BASEL; v. 11, n. 5, p. 27-pg., 2025-04-23. |
| Resumo | |
L-asparaginase (L-ASNase) is a key enzyme in the treatment of leukemia and lymphoma, with high demand in cancer therapies. Advances in recombinant protein production have improved yields and reduced costs, enabling large-scale production. However, optimizing culture conditions remains crucial for maximizing production. This study focused on optimizing the production of double mutant L-ASNase expressed in Escherichia coli BL21 (DE3) by supplementing media with amino acids. Five amino acids were evaluated at a shake flask scale using the design of experiments, with arginine and aspartate showing the most positive effects. Under optimized conditions (14.5 mM arginine, 12.7 mM aspartate, and 0 mM cysteine), the activity model reached 12,513 U L-1, experimentally validated at 10,089 U L-1. The maximum specific cell growth rate was mu x,max = 0.74 h-1, with substrate-biomass conversion factor Yx/s = 1.16 g/g and cell-product conversion factor Yp/x = 13,891 U/gcell. Amino acid supplementation resulted in a ten-fold increase in L-ASNase activity. Finally, at the bioreactor scale, adding amino acids and the inducer at the end of the exponential phase increased activity by 135% compared to conventional MD, demonstrating its potential for industrial-scale production. (AU) | |
| Processo FAPESP: | 20/06982-3 - L-asparaginase quimérica e peguilada: estudo das condições de expressão, produção e purificação de biofármaco antileucêmico inovador com potencial de baixa imunogenicidade |
| Beneficiário: | Adalberto Pessoa Junior |
| Modalidade de apoio: | Auxílio à Pesquisa - Regular |