HSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation
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| Publication Date: | 2014 |
| Other Authors: | , , , , , |
| Format: | Article |
| Language: | eng |
| Source: | Repositório Institucional da UnB |
| Download full: | http://repositorio.unb.br/handle/10482/22046 http://dx.doi.org/10.1590/S1517-83822014000200015 |
Summary: | 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. |
| _version_ | 1871442494272569344 |
|---|---|
| author | Zepeda, Andrea B. |
| author2 | Figueroa, Carolina A. Abdalla, Dulcineia Saes Parra Maranhão, Andrea Queiroz Ulloa, Patricio H. Pessoa Junior, Adalberto Farías, Jorge G. |
| author2_role | author author author author author author |
| author_browse | Abdalla, Dulcineia Saes Parra Farías, Jorge G. Figueroa, Carolina A. Maranhão, Andrea Queiroz Pessoa Junior, Adalberto Ulloa, Patricio H. Zepeda, Andrea B. |
| author_facet | Zepeda, Andrea B. Figueroa, Carolina A. Abdalla, Dulcineia Saes Parra Maranhão, Andrea Queiroz Ulloa, Patricio H. Pessoa Junior, Adalberto Farías, Jorge G. |
| author_role | author |
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| bitstream.url.fl_str_mv | http://repositorio2.unb.br/jspui/bitstream/10482/22046/1/ARTIGO_HSF-1%2c%20HIF-1and%20HSP90.pdf http://repositorio2.unb.br/jspui/bitstream/10482/22046/2/license.txt |
| collection | Repositório Institucional da UnB |
| dc.contributor.author.fl_str_mv | Zepeda, Andrea B. Figueroa, Carolina A. Abdalla, Dulcineia Saes Parra Maranhão, Andrea Queiroz Ulloa, Patricio H. Pessoa Junior, Adalberto Farías, Jorge G. |
| dc.date.accessioned.fl_str_mv | 2016-12-23T14:24:54Z |
| dc.date.available.fl_str_mv | 2016-12-23T14:24:54Z |
| dc.date.issued.fl_str_mv | 2014-04 |
| dc.identifier.citation.fl_str_mv | ZEPEDA, Andrea B. et al . HSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation. Brazilian Journal of Microbiology, São Paulo, v. 45, n. 2, p. 485-490, abr./jun. 2014. Disponível em: <http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000200015&lng=pt&nrm=iso>. Acessos em: 14 dez. 2016. http://dx.doi.org/10.1590/S1517-83822014000200015. |
| dc.identifier.doi.en.fl_str_mv | http://dx.doi.org/10.1590/S1517-83822014000200015 |
| dc.identifier.uri.fl_str_mv | http://repositorio.unb.br/handle/10482/22046 |
| dc.language.iso.fl_str_mv | eng |
| dc.publisher.none.fl_str_mv | Sociedade Brasileira de Microbiologia |
| dc.rights.driver.fl_str_mv | info:eu-repo/semantics/openAccess |
| dc.source.none.fl_str_mv | reponame:Repositório Institucional da UnB instname:Universidade de Brasília (UnB) instacron:UNB |
| dc.subject.keyword.en.fl_str_mv | Leveduras Temperatura - efeito fisiológico Hipóxia |
| dc.title.en.fl_str_mv | HSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation |
| dc.type.driver.fl_str_mv | info:eu-repo/semantics/article |
| dc.type.status.fl_str_mv | info:eu-repo/semantics/publishedVersion |
| 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. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | UNB_c8fd4bcb3ed55df4d86d96876a768c55 |
| identifier_str_mv | ZEPEDA, Andrea B. et al . HSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation. Brazilian Journal of Microbiology, São Paulo, v. 45, n. 2, p. 485-490, abr./jun. 2014. Disponível em: <http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000200015&lng=pt&nrm=iso>. Acessos em: 14 dez. 2016. http://dx.doi.org/10.1590/S1517-83822014000200015. |
| instacron_str | UNB |
| institution | UNB |
| instname_str | Universidade de Brasília (UnB) |
| language | eng |
| network_acronym_str | UNB |
| network_name_str | Repositório Institucional da UnB |
| oai_identifier_str | oai:repositorio.unb.br:10482/22046 |
| publishDate | 2014 |
| publishDateSort | 2014 |
| publisher.none.fl_str_mv | Sociedade Brasileira de Microbiologia |
| reponame_str | Repositório Institucional da UnB |
| repository.mail.fl_str_mv | repositorio@unb.br |
| repository.name.fl_str_mv | Repositório Institucional da UnB - Universidade de Brasília (UnB) |
| repository_id_str | |
| spelling | Zepeda, Andrea B.Figueroa, Carolina A.Abdalla, Dulcineia Saes ParraMaranhão, Andrea QueirozUlloa, Patricio H.Pessoa Junior, AdalbertoFarías, Jorge G.2016-12-23T14:24:54Z2016-12-23T14:24:54Z2014-04ZEPEDA, Andrea B. et al . HSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation. Brazilian Journal of Microbiology, São Paulo, v. 45, n. 2, p. 485-490, abr./jun. 2014. Disponível em: <http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000200015&lng=pt&nrm=iso>. Acessos em: 14 dez. 2016. http://dx.doi.org/10.1590/S1517-83822014000200015.http://repositorio.unb.br/handle/10482/22046http://dx.doi.org/10.1590/S1517-83822014000200015Sociedade Brasileira de MicrobiologiaBrazilian Journal of Microbiology - All the content of the journal, except where otherwise noted, is licensed under a Creative Commons License CC BY-NC. Fonte: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000200015&lng=pt&nrm=iso. Acesso em: 14 dez. 2016.info:eu-repo/semantics/openAccessHSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentationinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleLevedurasTemperatura - efeito fisiológicoHipóxiaPichia 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.engreponame:Repositório Institucional da UnBinstname:Universidade de Brasília (UnB)instacron:UNBORIGINALARTIGO_HSF-1, HIF-1and HSP90.pdfARTIGO_HSF-1, HIF-1and HSP90.pdfapplication/pdf1030048http://repositorio2.unb.br/jspui/bitstream/10482/22046/1/ARTIGO_HSF-1%2c%20HIF-1and%20HSP90.pdfccbba18e3e4f89a34b93d90a49245654MD51open accessLICENSElicense.txtlicense.txttext/plain253http://repositorio2.unb.br/jspui/bitstream/10482/22046/2/license.txt7ee8337f672551ca7e9c1175b8c4681eMD52open access10482/220462024-07-16 08:44:42.755open accessoai:repositorio.unb.br:10482/22046TGljZW5zZSBncmFudGVkIGJ5IENhbWlsYSAgRHVhcnRlIChjYW1pbGFkaWFzQGJjZS51bmIuYnIpIG9uIDIwMTYtMTItMTRUMTM6MTY6MDZaIChHTVQpOgoKU3VibWlzc8OjbyBlZmV0aXZhZGEgcG9yIGludGVncmFudGUgZGEgZXF1aXBlIGRvIFJlcG9zaXTDs3JpbyBJbnN0aXR1Y2lvbmFsIGRhIFVuQiBkZSBhY29yZG8gY29tIGxpY2Vuw6dhIGNvbmNlZGlkYSBwZWxvIGF1dG9yIGUvb3UgZGV0ZW50b3IgZG9zIGRpcmVpdG9zIGF1dG9yYWlzLg==Repositório InstitucionalPUBhttps://repositorio.unb.br/oai/requestrepositorio@unb.bropendoar:2024-07-16T11:44:42Repositório Institucional da UnB - Universidade de Brasília (UnB) |
| spellingShingle | HSF-1, HIF-1and HSP90 expression on recombinant Pichia pastoris under fed-batch fermentation Zepeda, Andrea B. Leveduras Temperatura - efeito fisiológico Hipóxia |
| status_str | publishedVersion |
| 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 | Leveduras Temperatura - efeito fisiológico Hipóxia |
| url | http://repositorio.unb.br/handle/10482/22046 http://dx.doi.org/10.1590/S1517-83822014000200015 |