Biochemical characterization ofa b-fructofuranosidase from Rhodotorula dairenensiswith transfructosylating activity

An extracellular b-fructofuranosidase from the yeast Rhodotorula dairenensis was characterized biochemically. The enzyme molecular mass was estimated to be 680 kDa by analytical gel filtration and 172 kDa by sodium dodecyl sulfatepolyacrylamide gel electrophoresis, of which the N-linked carbohydrate...

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Detalles Bibliográficos
Autores: Gutiérrez-Alonso, Patricia, Fernández Arrojo, Lucía, Plou Gasca, Francisco José, Fernández Lobato, María
Tipo de recurso: artículo
Fecha de publicación:2009
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/24892
Acceso en línea:http://hdl.handle.net/10261/24892
Access Level:acceso abierto
Palabra clave:Rhodotorula dairenensis
Fructofuranosidase
Invertase
fructosyltransferase
prebiotic oligosaccharides
Descripción
Sumario:An extracellular b-fructofuranosidase from the yeast Rhodotorula dairenensis was characterized biochemically. The enzyme molecular mass was estimated to be 680 kDa by analytical gel filtration and 172 kDa by sodium dodecyl sulfatepolyacrylamide gel electrophoresis, of which the N-linked carbohydrate accounts for 16% of the total mass. It displays optimum activity at pH 5 and 55–60 1C. The enzyme shows broad substrate specificity, hydrolyzing sucrose, 1-kestose, nystose, leucrose, raffinose and inulin. Although the main reaction catalyzed by this enzyme is sucrose hydrolysis, it also exhibits transfructosylating activity that, unlike other microbial b-fructofuranosidases, produces a varied type of prebiotic fructooligosaccharides containing b-(2 ! 1)- and b-(2 ! 6)-linked fructose oligomers. The maximum concentration of fructooligosaccharides was reached at 75% sucrose conversion and it was 87.9 g L!1. The 17.0% (w/w) referred to the total amount of sugars in the reaction mixture. At this point, the amounts of 6- kestose, neokestose, 1-kestose and tetrasaccharides were 68.9, 10.6, 2.6 and 12.7 g L!1, respectively.