Production of co-immobilized dextransucrase and dextranase preparations and their application in isomalto-oligosaccharides synthesis

Abstract

Dextransucrase (DS) from Leuconostoc mesenteroides and dextranase (DN) from Penicillium funiculosum were co-immobilized by entrapment in calcium alginate and used to produce isomaltooligosaccharides (IMOs) from sucrose. DS convert sucrose into dextran, which is the substrate for DN, so that IMOs are products of dextran hydrolysis. Before the co-immobilization DS was cross-linked with glutardialdehyde (GA), while DN was adsorbed on hydroxyapatite (HAp). Cross-linking was essential for the stability of DS and pre-immobilization of DN to prevent enzyme from leaking out of the alginate beads. Operational stability of co-immobilized preparations of DS and DN was estimated based on amounts of isomaltose and isomaltotriose formed during successive 24h processes of IMOs synthesis, carried out at 30oC, pH 5.4 and 200 rpm in 10% (w/v) sucrose solutions. Preparation characterized by the initial DS/DN activities ratio of 1/14 was found to maintain these activities at least 100 h of IMOs synthesis (5 repeated batch reaction).