Biomarkers to evaluate the effects of temperature and methanol on recombinant Pichia pastoris

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Autor(es): dc.creatorZepeda, Andrea B.-
Autor(es): dc.creatorFigueroa, Carolina A.-
Autor(es): dc.creatorAbdalla, Dulcineia Saes Parra-
Autor(es): dc.creatorMaranhão, Andrea Queiroz-
Autor(es): dc.creatorUlloa, Patricio H.-
Autor(es): dc.creatorPessoa Junior, Adalberto-
Autor(es): dc.creatorFarías, Jorge G.-
Data de aceite: dc.date.accessioned2024-07-22T12:24:20Z-
Data de disponibilização: dc.date.available2024-07-22T12:24:20Z-
Data de envio: dc.date.issued2016-12-21-
Data de envio: dc.date.issued2016-12-21-
Data de envio: dc.date.issued2014-04-
Fonte completa do material: dc.identifierhttp://repositorio.unb.br/handle/10482/22014-
Fonte completa do material: dc.identifierhttp://dx.doi.org/10.1590/S1517-83822014000200014-
Fonte: dc.identifier.urihttp://educapes.capes.gov.br/handle/capes/802948-
Descrição: dc.descriptionPichia 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 (GLR) and TBARS. In addition, there was a cytosolic expression of HSF-1 and HSP-70, which indicates a deprotection against nucleolar fragmentation (apoptosis). On the other hand, at 3% methanol -10 °C and 1% and at methanol -10 °C conditions there was nuclear expression of OGG1, lower levels of TBARS and lower expression of GLR, cytosolic expression of HSF-1 and nuclear expression HSP-70. In conclusion, our results suggest that 3% methanol-30 °C is a condition that induces a strong oxidative stress and risk factors of apoptosis in modified-genetically P. pastoris.-
Formato: dc.formatapplication/pdf-
Publicador: dc.publisherSociedade Brasileira de Microbiologia-
Direitos: dc.rightsAcesso Aberto-
Direitos: dc.rightsBrazilian 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=S1517-83822014000200014&lng=pt&nrm=iso. Acesso em: 14 dez. 2016.-
Palavras-chave: dc.subjectStress oxidativo-
Palavras-chave: dc.subjectLeveduras-
Palavras-chave: dc.subjectTemperatura - efeito fisiológico-
Palavras-chave: dc.subjectDNA-
Título: dc.titleBiomarkers to evaluate the effects of temperature and methanol on recombinant Pichia pastoris-
Aparece nas coleções:Repositório Institucional – UNB

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