HSF-1, HIF-1 and HSP90 expression on recombinant Pichia pas of is under fed-batch fermentation

Autor(es)

Zepeda, Andrea B. ^{[1, 2]} ; Figueroa, Carolina A. ^{[1, 2]} ; Abdalla, Dulcineia S. P. ^[3] ; Maranhao, Andrea Q. ^[4] ; Ulloa, Patricia H. ^[2] ; Pessoa, Jr., Adalberto ^[1] ; Farias, Jorge G. ^[2]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Fac Ciencias Farmaceut, Dept Tecnol Bioquim Farmaceut, Sao Paulo - Brazil

^[2] Univ La Frontera, Fac Ingn Ciencias & Adm, Dept Ingn Quim, Temuco - Chile

^[3] Univ Sao Paulo, Fac Ciencias Farmaceut, Dept Anal Clin & Toxicol, Sao Paulo - Brazil

^[4] Univ Brasilia, Inst Ciencias Biol, Dept Biol Celular, Brasilia, DF - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Brazilian Journal of Microbiology; v. 45, n. 2, p. 485-490, APR-JUN 2014.
• Citações Web of Science: 7

Resumo

Piehia pastoris is a methylotrophic yeast used as an efficient expression system for heterologous protein production as compared to other expression systems. Considering that every cell must respond to environmental changes to survive and differentiate, determination of endogenous protein related to heat stress responses and hypoxia, it would necessary to establish the temperature and methanol concentration conditions for optimal growth. The aim of this study is characterize the culture conditions through the putative biomarkers in different conditions of temperature and methanol concentration. Three yeast cultures were performed: 3X = 3% methanol -10 C, 4X = 3% methanol -30 C, and 5X 1% methanol -10 C. The expression level of HU.- I a, HSF-1, HSP-70 and HSP-90 biomarkers were measured by Western blot and in situ detection was performed by immunocytochemistry. The western blot results of H1F-la and HSP-90 did not indicate statistically significant in the culture conditions studied. Respect to biomarkers location, HIF-la and HSP-90 presented differences between cultures, In conclusion, the results suggest the cultures in a hypoxic condition produce a high density and yeast cells smaller, Beside the high density would not necessary related with a high production of recombinant proteins in modified-genetically P. pastor/s. (AU)

• Processo FAPESP: 12/50210-9 - Produção de fragmentos recombinantes de anticorpos a partir do cultivo de Pichia pastoris geneticamente modificada
• Beneficiário: Adalberto Pessoa Junior
• Modalidade de apoio: Auxílio à Pesquisa - Regular