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dc.contributor.authorZepeda, Andrea B.-
dc.contributor.authorFigueroa, Carolina A.-
dc.contributor.authorAbdalla, Dulcineia Saes Parra-
dc.contributor.authorMaranhão, Andrea Queiroz-
dc.contributor.authorUlloa, Patricio H.-
dc.contributor.authorPessoa Junior, Adalberto-
dc.contributor.authorFarías, Jorge G.-
dc.date.accessioned2016-12-21T12:01:53Z-
dc.date.available2016-12-21T12:01:53Z-
dc.date.issued2014-04-
dc.identifier.citationZEPEDA, Andrea B. et al. Biomarkers to evaluate the effects of temperature and methanol on recombinant Pichia pastoris. Brazilian Journal of Microbiology, São Paulo, v. 45, n. 2, p. 475-483, abr./jun. 2014. Disponível em: <http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000200014&lng=pt&nrm=iso>. Acesso em: 14 dez. 2016. http://dx.doi.org/10.1590/S1517-83822014000200014.en
dc.identifier.urihttp://repositorio.unb.br/handle/10482/22014-
dc.description.abstractPichia 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.en
dc.language.isoInglêsen
dc.publisherSociedade Brasileira de Microbiologiaen
dc.rightsAcesso Abertoen
dc.titleBiomarkers to evaluate the effects of temperature and methanol on recombinant Pichia pastorisen
dc.typeArtigoen
dc.subject.keywordStress oxidativoen
dc.subject.keywordLevedurasen
dc.subject.keywordTemperatura - efeito fisiológicoen
dc.subject.keywordDNAen
dc.rights.licenseBrazilian 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.en
dc.identifier.doihttp://dx.doi.org/10.1590/S1517-83822014000200014en
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