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Biomarkers to evaluate the effects of temperature and methanol on recombinant Pichia pastoris BJM
Zepeda,Andrea B.; Figueroa,Carolina A.; Abdalla,Dulcineia S.P.; Maranhão,Andrea Q.; Ulloa,Patricio H.; Pessoa Jr,Adalberto; Farías,Jorge G..
Pichia pastoris is methylotrophic yeast used as an efficient expression system for heterologous protein production. In order to evaluate the effects of temperature (10 and 30 °C) and methanol (1 and 3% (v/v)) on genetically-modified Pichia pastoris, different biomarkers were evaluated: Heat stress (HSF-1 and Hsp70), oxidative stress (OGG1 and TBARS) and antioxidant (GLR). 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. Ours results show that at 3% methanol -30 °C there is an increase of mitochondrial OGG1 (mtOGG1), Glutathione Reductase...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Oxidative stress; Recombinant yeast; Heat stress protein; DNA damage.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000200014
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HSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation BJM
Zepeda,Andrea B.; Figueroa,Carolina A.; Abdalla,Dulcineia S.P.; Maranhão,Andrea Q.; Ulloa,Patricio H.; Pessoa Jr.,Adalberto; Farías,Jorge G..
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...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Recombinant yeast; Heat stress protein; Hypoxia; Chaperone.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000200015
Registros recuperados: 2
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