Details
| Originalsprache | Englisch |
|---|---|
| Seiten (von - bis) | 49-56 |
| Seitenumfang | 8 |
| Fachzeitschrift | Biocatalysis and biotransformation |
| Jahrgang | 22 |
| Ausgabenummer | 1 |
| Publikationsstatus | Veröffentlicht - Jan. 2004 |
Abstract
GDP-β-L-fucose synthetase (GFS) is a unique, bifunctional enzyme belonging to the family of short chain dehydrogenases (SDRs), which catalyses the conversion of GDP-4-keto-6-deoxy-α-D-mannose (GKDM) to GDP-β-L-fucose (GDP-Fuc). A facile method for preparing GKDM in a stable and storable form on a 200 μmol scale was established. A simple macroscopic kinetic model has been used to produce a tool for simulation of several GFS reactions in batch mode. The respective kinetic parameters were determined considering GFS using NADPH and NADH as cofactor, demonstrating that GFS has a different affinity for GKDM depending on the cofactor. A comparison of simulated and experimentally performed batch reactions revealed that the GFS reaction can be described by this simple kinetic model that takes into account product inhibition by GDP-Fuc and NADP or NAD. Full 1H- and 13C-NMR characterisation of GKDM unambiguously showed that this compound exists in two different forms in aqueous solution, which we identified as the ketone and the respective hydrate form of GKDM. In equilibrium these species coexist in a ratio of about 70:30 (keto/hydrate form).
ASJC Scopus Sachgebiete
- Biochemie, Genetik und Molekularbiologie (insg.)
- Biotechnologie
- Chemische Verfahrenstechnik (insg.)
- Katalyse
- Biochemie, Genetik und Molekularbiologie (insg.)
- Biochemie
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in: Biocatalysis and biotransformation, Jahrgang 22, Nr. 1, 01.2004, S. 49-56.
Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review
}
TY - JOUR
T1 - Kinetic examination and simulation of GDP-β-L-fucose synthetase reaction using NADPH or NADH
AU - Rentmeister, Andrea
AU - Hoh, Christoph
AU - Weidner, Stefan
AU - Dräger, Gerald
AU - Elling, Lothar
AU - Liese, Andreas
AU - Wandrey, Christian
N1 - Copyright: Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2004/1
Y1 - 2004/1
N2 - GDP-β-L-fucose synthetase (GFS) is a unique, bifunctional enzyme belonging to the family of short chain dehydrogenases (SDRs), which catalyses the conversion of GDP-4-keto-6-deoxy-α-D-mannose (GKDM) to GDP-β-L-fucose (GDP-Fuc). A facile method for preparing GKDM in a stable and storable form on a 200 μmol scale was established. A simple macroscopic kinetic model has been used to produce a tool for simulation of several GFS reactions in batch mode. The respective kinetic parameters were determined considering GFS using NADPH and NADH as cofactor, demonstrating that GFS has a different affinity for GKDM depending on the cofactor. A comparison of simulated and experimentally performed batch reactions revealed that the GFS reaction can be described by this simple kinetic model that takes into account product inhibition by GDP-Fuc and NADP or NAD. Full 1H- and 13C-NMR characterisation of GKDM unambiguously showed that this compound exists in two different forms in aqueous solution, which we identified as the ketone and the respective hydrate form of GKDM. In equilibrium these species coexist in a ratio of about 70:30 (keto/hydrate form).
AB - GDP-β-L-fucose synthetase (GFS) is a unique, bifunctional enzyme belonging to the family of short chain dehydrogenases (SDRs), which catalyses the conversion of GDP-4-keto-6-deoxy-α-D-mannose (GKDM) to GDP-β-L-fucose (GDP-Fuc). A facile method for preparing GKDM in a stable and storable form on a 200 μmol scale was established. A simple macroscopic kinetic model has been used to produce a tool for simulation of several GFS reactions in batch mode. The respective kinetic parameters were determined considering GFS using NADPH and NADH as cofactor, demonstrating that GFS has a different affinity for GKDM depending on the cofactor. A comparison of simulated and experimentally performed batch reactions revealed that the GFS reaction can be described by this simple kinetic model that takes into account product inhibition by GDP-Fuc and NADP or NAD. Full 1H- and 13C-NMR characterisation of GKDM unambiguously showed that this compound exists in two different forms in aqueous solution, which we identified as the ketone and the respective hydrate form of GKDM. In equilibrium these species coexist in a ratio of about 70:30 (keto/hydrate form).
KW - Enzymatic synthesis
KW - GDP-α-D-mannose-4,6-dehydratase
KW - GDP-β-L-fucose synthethase
KW - GDP-4-keto-6-deoxy-α-D-mannose
KW - Kinetic examination and simulation
KW - Nicotinamide cofactors
UR - http://www.scopus.com/inward/record.url?scp=1842588372&partnerID=8YFLogxK
U2 - 10.1080/10242420410001666362
DO - 10.1080/10242420410001666362
M3 - Article
AN - SCOPUS:1842588372
VL - 22
SP - 49
EP - 56
JO - Biocatalysis and biotransformation
JF - Biocatalysis and biotransformation
SN - 1024-2422
IS - 1
ER -