Artículo
Optimization of the Production of a Polygalacturonase from Aspergillus kawachii cloned in Saccharomyces cerevisiae in batch and fed-batch cultures
Rojas, Natalia Lorena
; Ortiz, Gastón Ezequiel
; Chesini, Mariana
; Baruque, Diego Jorge; Cavalitto, Sebastian Fernando
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Fecha de publicación:
02/2011
Editorial:
University of Zagreb. Faculty of Food Technology and Biotechnology
Revista:
Food Technology and Biotechnology
ISSN:
1330-9862
e-ISSN:
1334-2606
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
Polygalacturonases (PG; EC 3.2.1.15) catalyze the hydrolysis of pectin and/or pectic acid and are useful for industrial applications such as juice clarification and pectin extraction. Growth and heterologous expression of recombinant S. cerevisiae which expresses an acidic PG from Aspergillus kawachii was studied in batch and fed batch cultures. Kinetics and stoichiometric parameters of the recombinant yeast were determined in batch cultures in a synthetic medium. In these cultures, the total biomass concentration, protein concentration, and enzyme activity achieved were 2.2 g/l, 10 mg/l, and 3 U/ml, respectively, to give a productivity of 0.06 U/ml.h. In fed batch cultures, various strategies for galactose feeding were used: a) after a glucose growth phase, the addition of a single pulse of galactose which gave a productivity of 0.19 U/ml.h.; b) after a glucose growth phase, a double pulse of galactose at the same final concentration, resulting in a productivity of 0.21 U/ml.h.; c) a simultaneous feeding of glucose and galactose, yielding a productivity of 1.32 U/ml.h. Based on these results, the simultaneous feeding of glucose and galactose was by far the most suitable strategy for the production of this enzyme. Moreover, some biochemical characteristics of the recombinant enzyme such as a MW of ~60 kDa, an isoelectric point of 3.7 and its ability to hydrolyze polygalacturonic acid at pH 2.5 were determined.
Palabras clave:
PG1
Archivos asociados
Licencia
Identificadores
Colecciones
Articulos(CINDEFI)
Articulos de CENT.DE INV EN FERMENTACIONES INDUSTRIALES (I)
Articulos de CENT.DE INV EN FERMENTACIONES INDUSTRIALES (I)
Articulos(IBYME)
Articulos de INST.DE BIOLOGIA Y MEDICINA EXPERIMENTAL (I)
Articulos de INST.DE BIOLOGIA Y MEDICINA EXPERIMENTAL (I)
Citación
Rojas, Natalia Lorena; Ortiz, Gastón Ezequiel; Chesini, Mariana; Baruque, Diego Jorge; Cavalitto, Sebastian Fernando; Optimization of the Production of a Polygalacturonase from Aspergillus kawachii cloned in Saccharomyces cerevisiae in batch and fed-batch cultures; University of Zagreb. Faculty of Food Technology and Biotechnology; Food Technology and Biotechnology; 49; 2-2011; 316-321
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