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Toward Automated Enzymatic Glycan Synthesis in a Compartmented Flow Microreactor System

Heinzler, Raphael; Fischöder, T.; Elling, L.; Franzreb, Matthias

Abstract:
Immobilized microfluidic enzyme reactors (IMER) are of particular interest for automation of enzyme cascade reactions. Within an IMER, substrates are converted by paralleled immobilized enzyme modules and intermediate products are transported for further conversion by subsequent enzyme modules. By optimizing substrate conversion in the spatially separated enzyme modules purification of intermediate products is not necessary, thus shortening process time and increasing space‐time yields. The IMER enables the development of efficient enzyme cascades by combining compatible enzymatic reactions in different arrangements under optimal conditions and the possibility of a cost‐benefit analysis prior to scale‐up. These features are of special interest for automation of enzymatic glycan synthesis. We here demonstrate a compartmented flow microreactor system using six magnetic enzyme beads (MEBs) for the synthesis of the non‐sulfated human natural killer cell‐1 (HNK‐1) glycan epitope. MEBs are assembled to build compartmented enzyme modules, consisting of enzyme cascades for the synthesis of uridine 5′‐ diphospho‐α‐ d‐galactose (UDP‐Gal) and uridine 5′‐diphospho‐α‐d‐glucuronic acid (UDP‐GlcA), the donor substrates for the Leloir glycosyltransferases β4‐galactosyltransferase and β3‐glucuronosyltransferase, respectively. ... mehr

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Verlagsausgabe §
DOI: 10.5445/IR/1000098147
Veröffentlicht am 13.09.2019
Coverbild
Zugehörige Institution(en) am KIT Institut für Funktionelle Grenzflächen (IFG)
Publikationstyp Zeitschriftenaufsatz
Jahr 2019
Sprache Englisch
Identifikator ISSN: 0941-1216, 1436-9966, 1521-3897, 1615-4150, 1615-4169
KITopen-ID: 1000098147
HGF-Programm 47.02.06 (POF III, LK 01)
Erschienen in Advanced synthesis & catalysis
Band 361
Heft 19
Seiten 4506-4516
Vorab online veröffentlicht am 21.07.2019
Schlagworte Glycoconjugate; Microreactors; Biocatalysis; Immobilization; Magnetic beads
Nachgewiesen in Scopus
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