title: Immobilization and Characterization of E. gracilis Extract with Enriched Laminaribiose Phosphorylase Activity for Bienzymatic Production of Laminaribiose.
abstract: Immobilization methods and carriers were screened for immobilization of Euglena gracilis extract with laminaribiose phosphorylase activity. The extract was successfully immobilized on three different carriers via covalent linkage. Suitable immobilization carriers were Sepabeads EC-EP/S and ECR 8209M with epoxy groups and ECR 8309M with amino groups as functional units. Immobilization on Sepabeads EC-EP/S resulted in highest retained activity (65%). The immobilizates were characterized for pH, temperature, and buffer molarity preferences. The immobilized enzyme lost 48% of its activity when used seven times. Together with sucrose phosphorylase, laminaribiose phosphorylase was successfully applied for bienzymatic production of laminaribiose from sucrose and glucose with a final laminaribiose concentration of 14.3 ± 2.1 g/L (20% yield).
authors.affiliation: TU Braunschweig, Braunschweig, Germany.
authors.forename: Clarissa
authors.initials: C
authors.lastname: Müller
authors.affiliation: TU Braunschweig, Braunschweig, Germany.
authors.forename: Tim
authors.initials: T
authors.lastname: Ortmann
authors.affiliation: TU Braunschweig, Braunschweig, Germany.
authors.forename: Akram
authors.initials: A
authors.lastname: Abi
authors.affiliation: TU Braunschweig, Braunschweig, Germany.
authors.forename: Dave
authors.initials: D
authors.lastname: Hartig
authors.affiliation: TU Braunschweig, Braunschweig, Germany.
authors.forename: Stephan
authors.initials: S
authors.lastname: Scholl
authors.affiliation: TU Braunschweig, Braunschweig, Germany. A.Joerdening@tu-bs.de.
authors.forename: Hans-Joachim
authors.initials: HJ
authors.lastname: Jördening
mesh_terms: D002021:Buffers; D004187:Disaccharides / Q000138:chemical synthesis*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000302:isolation & purification; D004853:Epoxy Resins / Q000737:chemistry; D005056:Euglena gracilis / Q000737:chemistry / Q000201:enzymology*; D005163:Factor Analysis, Statistical; D005947:Glucose / Q000737:chemistry; D005964:Glucosyltransferases / Q000737:chemistry* / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D015800:Protozoan Proteins / Q000737:chemistry* / Q000302:isolation & purification; D013395:Sucrose / Q000737:chemistry; D013696:Temperature
chemical_list: D002021:Buffers; D004187:Disaccharides; D004800:Enzymes, Immobilized; D004853:Epoxy Resins; D015800:Protozoan Proteins; C014637:laminaribiose; D013395:Sucrose; D005964:Glucosyltransferases; C082478:laminaribiose phosphorylase; C025901:sucrose phosphorylase; D005947:Glucose
keywords: Bienzymatic system; Enzyme immobilization; Laminaribiose; Laminaribiose phosphorylase; Response surface methodology