Publicação
Novos transportadores de polióis em Debaryomyces hansenii
| Resumo: | Bioethanol production from lignocellulosic residues is a promising alternative for fossil fuels. However, it is still economically unviable due to the incapability of the main fermentative yeast, Saccharomyces cerevisiae, to ferment pentoses from hemicelluloses. In this work, a molecular approach was followed, as a contribution to construct genetically modified strains of S. cerevisiae expressing putative polyol transporters, in order to upgrade the hemicelluloses fermentation by producing simultaneously plus-valuable dietetical polyols for food industry, such as xylitol. A bibliographic screening for yeast species able to assimilate polyols revealed a restricted number of ascomycetes among the genus Candida, Debaryomyces and Pichia with potential interest as sinks of useful genes to improve polyol release. To identify polyol transporters, a BlastP search on D. hansenii CBS 767 genome was performed, followed by cloning and characterization in S. cerevisiae. The presence of polyol(sugar)/H+ symport activity was evaluated for 18 substrates based on the external alkalinization of unbuffered cell suspensions upon polyol(sugar) addition. In the present work 4 ORFs were cloned and 10 were characterized. Polyol/H+ symport activity (glycerol/H+, galactitol/H+, sorbitol/mannitol/ribitol/arabitol/H+) was detected in some of the strains tested. |
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| Autores principais: | Pereira, Iliana Bento |
| Assunto: | lignocellulosic waste bioethanol polyols debaryomyces hansenii |
| Ano: | 2011 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | português |
| Origem: | Repositório da Universidade de Lisboa |
| Resumo: | Bioethanol production from lignocellulosic residues is a promising alternative for fossil fuels. However, it is still economically unviable due to the incapability of the main fermentative yeast, Saccharomyces cerevisiae, to ferment pentoses from hemicelluloses. In this work, a molecular approach was followed, as a contribution to construct genetically modified strains of S. cerevisiae expressing putative polyol transporters, in order to upgrade the hemicelluloses fermentation by producing simultaneously plus-valuable dietetical polyols for food industry, such as xylitol. A bibliographic screening for yeast species able to assimilate polyols revealed a restricted number of ascomycetes among the genus Candida, Debaryomyces and Pichia with potential interest as sinks of useful genes to improve polyol release. To identify polyol transporters, a BlastP search on D. hansenii CBS 767 genome was performed, followed by cloning and characterization in S. cerevisiae. The presence of polyol(sugar)/H+ symport activity was evaluated for 18 substrates based on the external alkalinization of unbuffered cell suspensions upon polyol(sugar) addition. In the present work 4 ORFs were cloned and 10 were characterized. Polyol/H+ symport activity (glycerol/H+, galactitol/H+, sorbitol/mannitol/ribitol/arabitol/H+) was detected in some of the strains tested. |
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