Abstract
The pathway of l-arabinose degradation was studied in the haloarchaeon Haloferax volcanii. It is shown that l-arabinose is oxidatively degraded to α-ketoglutarate. During growth on l-arabinose, l-arabinose dehydrogenase (l-AraDH) was induced. The enzyme was purified as a 130 kDa homotetrameric protein catalyzing the oxidation of l-arabinose with both NADP+ and NAD+. The gene encoding l-AraDH was identified as HVO_B0032 and recombinant l-AraDH showed similar properties as the native enzyme. The l-AraDH deletion mutant did not grow on l-arabinose, but grew unaffected on glucose and d-xylose, indicating a specific involvement in l-arabinose degradation. Phylogenetic analyses attribute the first archaeal l-AraDH to the extended short-chain dehydrogenase/reductase (SDRe) family, where it is part of a novel cluster and thus differs from known archaeal and bacterial pentose dehydrogenases. Further, cell extracts of H. volcanii catalyzed the NADP+-dependent conversion of l-arabinoate to α-ketoglutarate. The genes involved in that conversion were identified by analyses of transcripts and deletion mutants as HVO_B0038A, HVO_B0027 and HVO_B0039 recently reported to be involved in d-xylonate conversion to α-ketoglutarate in H. volcanii (Johnsen et al. 2009).
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Acknowledgments
We thank Roland Schmid (Osnabrück, Germany) for performing N-terminal amino acid sequencing. Thanks are also due to Jörg Soppa (Frankfurt, Germany) for providing the plasmid pSD1 and for supporting to establish the method for 5′-end and 3′-end determination of transcripts in our lab. This work was supported by grants of the Deutsche Forschungsgemeinschaft (SCHO 316/11-1).
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Communicated by L. Huang.
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Johnsen, U., Sutter, JM., Zaiß, H. et al. l-Arabinose degradation pathway in the haloarchaeon Haloferax volcanii involves a novel type of l-arabinose dehydrogenase. Extremophiles 17, 897–909 (2013). https://doi.org/10.1007/s00792-013-0572-2
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DOI: https://doi.org/10.1007/s00792-013-0572-2