title: HSF-1, HIF-1 and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation.
abstract: Pichia 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 HIF-1α, 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 HIF-1α and HSP-90 did not indicate statistically significant in the culture conditions studied. Respect to biomarkers location, HIF-1α 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. pastoris.
authors.affiliation: Departamento de Ingeniería Química Facultad de Ingeniería, Ciencias y Administración Universidad de La Frontera Temuco Chile Departamento de Ingeniería Química, Facultad de Ingeniería, Ciencias y Administración, Universidad de La Frontera, Temuco, Chile. ; Departamento de Tecnologia Bioquímico-Farmacêutica Faculdade de Ciências Farmacêuticas Universidade de São Paulo São PauloSP Brazil Departamento de Tecnologia Bioquímico-Farmacêutica, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Andrea B
authors.initials: AB
authors.lastname: Zepeda
authors.affiliation: Departamento de Ingeniería Química Facultad de Ingeniería, Ciencias y Administración Universidad de La Frontera Temuco Chile Departamento de Ingeniería Química, Facultad de Ingeniería, Ciencias y Administración, Universidad de La Frontera, Temuco, Chile. ; Departamento de Tecnologia Bioquímico-Farmacêutica Faculdade de Ciências Farmacêuticas Universidade de São Paulo São PauloSP Brazil Departamento de Tecnologia Bioquímico-Farmacêutica, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Carolina A
authors.initials: CA
authors.lastname: Figueroa
authors.affiliation: Departamento de Análises Clínicas e Toxicológicas Faculdade de Ciências Farmacêuticas Universidade de São Paulo São PauloSP Brazil Departamento de Análises Clínicas e Toxicológicas, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Dulcineia S P
authors.initials: DS
authors.lastname: Abdalla
authors.affiliation: Departamento de Biología Celular Instituto de Ciências Biológicas Universidade de Brasília BrasíliaDF Brazil Departamento de Biología Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, Brazil.
authors.forename: Andrea Q
authors.initials: AQ
authors.lastname: Maranhão
authors.affiliation: Departamento de Ingeniería Química Facultad de Ingeniería, Ciencias y Administración Universidad de La Frontera Temuco Chile Departamento de Ingeniería Química, Facultad de Ingeniería, Ciencias y Administración, Universidad de La Frontera, Temuco, Chile.
authors.forename: Patricio H
authors.initials: PH
authors.lastname: Ulloa
authors.affiliation: Departamento de Tecnologia Bioquímico-Farmacêutica Faculdade de Ciências Farmacêuticas Universidade de São Paulo São PauloSP Brazil Departamento de Tecnologia Bioquímico-Farmacêutica, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Adalberto
authors.initials: A
authors.lastname: Pessoa
authors.affiliation: Departamento de Ingeniería Química Facultad de Ingeniería, Ciencias y Administración Universidad de La Frontera Temuco Chile Departamento de Ingeniería Química, Facultad de Ingeniería, Ciencias y Administración, Universidad de La Frontera, Temuco, Chile.
authors.forename: Jorge G
authors.initials: JG
authors.lastname: Farías
mesh_terms: D000693:Anaerobiosis; D061249:Batch Cell Culture Techniques; D015153:Blotting, Western; D005285:Fermentation; D005656:Fungal Proteins / Q000032:analysis*; D007150:Immunohistochemistry; D000432:Methanol / Q000378:metabolism; D010843:Pichia / Q000737:chemistry* / Q000254:growth & development*; D013696:Temperature
chemical_list: D005656:Fungal Proteins; D000432:Methanol
keywords: chaperone; heat stress protein; hypoxia; recombinant yeast