title: Evaluation of a high temperature immobilised enzyme reactor for production of non-reducing oligosaccharides.
abstract: There is interest in the production of non-reducing carbohydrates due to their potential application in various industrial fields, particularly the food industry. In this paper, we describe the development of an immobilised cell bioprocess for the synthesis of non-reducing maltodextrins at high temperatures. The trehalosyl-dextrins-forming enzyme (TDFE) isolated from the thermoacidophilic archaeon Sulfolobus solfataricus (strain MT4), was recently expressed at high yields in Escherichia coli (strain Rb-791). Here, we evaluate different matrices, such as polyacrylamide gel, crude egg white, chitosan and calcium alginate for their effectiveness in immobilising whole recombinant E. coli cells subjected to prior thermal permeabilisation. Calcium-alginate based gels formed a solid biocatalyst with a good activity yield and the best enzymatic stability at the operating temperature (75 degrees C). Therefore, these beads were used to pack a glass column reactor to perform the bioconversion of interest. Optimal operating parameters were defined in relation to the substrate stream flow-rate and the substrate-to-biocatalyst ratio. The production of trehalosylmaltotetraose from maltohexaose reached equilibrium with a constant of about 2.6 at 75 degrees C. The bioreactor was exploited for production of trehalosylmaltodextrins from a commercial mixture of maltodextrins, achieving a productivity of 106.5 mg ml(-1) h(-1) (g biocatalyst)(-1) with ~40% conversion when using a 30% (w/v) solution.
authors.affiliation: Department of Experimental Medicine, Section of Biotechnology and Molecular Biology, Second University of Naples, via De Crecchio n degrees 7, 80138, Naples, Italy. chiara.schiraldi@unina2.it
authors.forename: Chiara
authors.initials: C
authors.lastname: Schiraldi
authors.forename: Isabella
authors.initials: I
authors.lastname: Di Lernia
authors.forename: Mariateresa
authors.initials: M
authors.lastname: Giuliano
authors.forename: Maddalena
authors.initials: M
authors.lastname: Generoso
authors.forename: Antonella
authors.initials: A
authors.lastname: D'Agostino
authors.forename: Mario
authors.initials: M
authors.lastname: De Rosa
mesh_terms: D019149:Bioreactors; D001709:Biotechnology / Q000379:methods; D001711:Biotransformation; D018914:Cells, Immobilized / Q000378:metabolism; D003912:Dextrins / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D009844:Oligosaccharides / Q000032:analysis / Q000378:metabolism*; D030781:Organisms, Genetically Modified / Q000378:metabolism; D016944:Sulfolobus / Q000201:enzymology / Q000235:genetics
chemical_list: D003912:Dextrins; D009844:Oligosaccharides; C016549:maltohexaose