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| DOI | 10.4014/JMB.1805.05056 | ||||
| Año | 2018 | ||||
| Tipo | artículo de investigación |
Citas Totales
Autores Afiliación Chile
Instituciones Chile
% Participación
Internacional
Autores
Afiliación Extranjera
Instituciones
Extranjeras
Alginate lyases (endo and exo-lyases) are required for the degradation of alginate into its constituting monomers. Efficient bioethanol production and extraction of bioactives from brown algae requires intensive use of these enzymes. Nonetheless, there are few commercial alginate lyase preparations, and their costs make them unsuitable for large scale experiments. A recombinant expression protocol has been developed in this study for producing seven endo-lyases and three exo-lyases as soluble and highly active preparations. Saccharification of alginate using 21 different endo/exo-lyase combinations shows that there is complementary enzymatic activity between some of the endo/exo pairs. This is probably due to favorable matching of their substrate biases for the different glycosidic bonds in the alginate molecule. Therefore, selection of enzymes for the best saccharification results for a given biomass should be based on screens comprising both types of lyases. Additionally, different incubation temperatures, enzyme load ratios, and enzyme loading strategies were assessed using the best four enzyme combinations for treating Macrocystis pyrifera biomass. It was shown that 30 degrees C with a 1:3 endo/exo loading ratio was suitable for all four combinations. Moreover, simultaneous loading of endo-and exo-lyases at the beginning of the reaction allowed maximum alginate saccharification in half the time than when the exo-lyases were added sequentially.
| Ord. | Autor | Género | Institución - País |
|---|---|---|---|
| 1 | GIMPEL-HENRIQUEZ, JAVIER ANDRES | Hombre |
Universidad de Chile - Chile
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| 2 | Ravanal, Maria-Cristina | Mujer |
Universidad de Chile - Chile
Universidad Austral de Chile - Chile |
| 3 | SALAZAR-GUERRERO, OSVALDO | Mujer |
Universidad de Chile - Chile
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| 4 | LIENQUEO-CONTRERAS, MARIA ELENA | Mujer |
Universidad de Chile - Chile
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| 4 | Elena Lienqueo, Ma. | Mujer |
Universidad de Chile - Chile
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| Agradecimiento |
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| This work was supported by Comision Nacional de InvestigacionCientifica y Tecnologica CONICYT, Chile: [Grant AKA-ERNC 009 PROJECT] and [Centre for Biotechnology and Bioengineering, CEBIB FB-0001]. Our thanks to Dr. Julia Rossmanith, Dr. Hueseyin Besir, Dr. Arie Geerlof, and the Protein Expression and Purification Facility, EMBL, Heidelberg, for facilitating the Gateway expression vectors utilized in this study. |