Free fatty acids reduce metabolic stress and favor a stable production of heterologous proteins in Pichia pastoris

Autor(es)

Zepeda, Andrea B. ^{[1, 2]} ; Figueroa, Carolina A. ^{[1, 2]} ; Pessoa, Adalberto ^[2] ; Farias, Jorge G. ^[1]

Afiliação do(s) autor(es)

^[1] Univ La Frontera, Fac Ingn Ciencias & Adm, Dept Ingn Quim, Temuco - Chile

^[2] Univ Sao Paulo, Fac Ciencias Farmaceut, Dept Tecnol Bioquim Farmaceut, Sao Paulo, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Brazilian Journal of Microbiology; v. 49, n. 4, p. 856-864, OCT-DEC 2018.
• Citações Web of Science: 0

Resumo

ABSTRACT The growth of yeasts in culture media can be affected by many factors. For example, methanol can be metabolized by other pathways to produce ethanol, which acts as an inhibitor of the heterologous protein production pathway; oxygen concentration can generate aerobic or anaerobic environments and affects the fermentation rate; and temperature affects the central carbon metabolism and stress response protein folding. The main goal of this study was determine the implication of free fatty acids on the production of heterologous proteins in different culture conditions in cultures of Pichia pastoris. We evaluated cell viability using propidium iodide by flow cytometry and thiobarbituric acid reactive substances to measure cell membrane damage. The results indicate that the use of low temperatures and low methanol concentrations favors the decrease in lipid peroxidation in the transition phase from glycerol to methanol. In addition, a temperature of 14 ºC + 1%M provided the most stable viability. By contrast, the temperature of 18 ºC + 1.5%M favored the production of a higher antibody fragment concentration. In summary, these results demonstrate that the decrease in lipid peroxidation is related to an increased production of free fatty acids. (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