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Gewählte Publikation:

Zolghadr, B; Gasselhuber, B; Windwarder, M; Pabst, M; Kracher, D; Kerndl, M; Zayni, S; Hofinger-Horvath, A; Ludwig, R; Haltrich, D; Oostenbrink, C; Obinger, C; Kosma, P; Messner, P; Schaffer, C.
(2015): UDP-sulfoquinovose formation by Sulfolobus acidocaldarius
EXTREMOPHILES. 2015; 19(2): 451-467. FullText FullText_BOKU

Abstract:
The UDP-sulfoquinovose synthase Agl3 from Sulfolobus acidocaldarius converts UDP-d-glucose and sulfite to UDP-sulfoquinovose, the activated form of sulfoquinovose required for its incorporation into glycoconjugates. Based on the amino acid sequence, Agl3 belongs to the short-chain dehydrogenase/reductase enzyme superfamily, together with SQD1 from Arabidopsis thaliana, the only UDP-sulfoquinovose synthase with known crystal structure. By comparison of sequence and structure of Agl3 and SQD1, putative catalytic amino acids of Agl3 were selected for mutational analysis. The obtained data suggest for Agl3 a modified dehydratase reaction mechanism. We propose that in vitro biosynthesis of UDP-sulfoquinovose occurs through an NAD(+)-dependent oxidation/dehydration/enolization/sulfite addition process. In the absence of a sulfur donor, UDP-d-glucose is converted via UDP-4-keto-d-glucose to UDP-d-glucose-5,6-ene, the structure of which was determined by H-1 and C-13-NMR spectroscopy. During the redox reaction the cofactor remains tightly bound to Agl3 and participates in the reaction in a concentration-dependent manner. For the first time, the rapid initial electron transfer between UDP-d-glucose and NAD(+) could be monitored in a UDP-sulfoquinovose synthase. Deuterium labeling confirmed that dehydration of UDP-d-glucose occurs only from the enol form of UDP-4-keto-glucose. The obtained functional data are compared with those from other UDP-sulfoquinovose synthases. A divergent evolution of Agl3 from S. acidocaldarius is suggested.
Autor/innen der BOKU Wien:
Gasselhuber Bernhard
Haltrich Dietmar
Hofinger-Horvath Andreas
Kosma Paul
Kracher Daniel
Ludwig Roland
Messner Paul
Obinger Christian
Oostenbrink Chris
Pabst Martin
Schäffer Christina
Windwarder Markus
Zayni Sonja
Zolghadr Behnam
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Find related publications in this database (Keywords)
UDP-sulfoquinovose synthase Agl3
Sulfolobus acidocaldarius
Enzyme mechanism
Site-directed mutagenesis
Sulfite


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