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HSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation

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...

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Autores principales: Zepeda, Andrea B., Figueroa, Carolina A., Abdalla, Dulcineia S.P., Maranhão, Andrea Q., Ulloa, Patricio H., Pessoa, Adalberto, Farías, Jorge G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Sociedade Brasileira de Microbiologia 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4166272/
https://www.ncbi.nlm.nih.gov/pubmed/25242931
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author Zepeda, Andrea B.
Figueroa, Carolina A.
Abdalla, Dulcineia S.P.
Maranhão, Andrea Q.
Ulloa, Patricio H.
Pessoa, Adalberto
Farías, Jorge G.
author_facet Zepeda, Andrea B.
Figueroa, Carolina A.
Abdalla, Dulcineia S.P.
Maranhão, Andrea Q.
Ulloa, Patricio H.
Pessoa, Adalberto
Farías, Jorge G.
author_sort Zepeda, Andrea B.
collection PubMed
description 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.
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spelling pubmed-41662722014-09-29 HSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation Zepeda, Andrea B. Figueroa, Carolina A. Abdalla, Dulcineia S.P. Maranhão, Andrea Q. Ulloa, Patricio H. Pessoa, Adalberto Farías, Jorge G. Braz J Microbiol Industrial Microbiology 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. Sociedade Brasileira de Microbiologia 2014-08-29 /pmc/articles/PMC4166272/ /pubmed/25242931 Text en Copyright © 2014, Sociedade Brasileira de Microbiologia All the content of the journal, except where otherwise noted, is licensed under a Creative Commons License CC BY-NC.
spellingShingle Industrial Microbiology
Zepeda, Andrea B.
Figueroa, Carolina A.
Abdalla, Dulcineia S.P.
Maranhão, Andrea Q.
Ulloa, Patricio H.
Pessoa, Adalberto
Farías, Jorge G.
HSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation
title HSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation
title_full HSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation
title_fullStr HSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation
title_full_unstemmed HSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation
title_short HSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation
title_sort hsf-1, hif-1and hsp90 expression on recombinant pichia pastoris under fed-batch fermentation
topic Industrial Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4166272/
https://www.ncbi.nlm.nih.gov/pubmed/25242931
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