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A novel trehalose synthesizing pathway in the hyperthermophilic Crenarchaeon Thermoproteus tenax: the unidirectional TreT pathway

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Abstract

In the genome of the hyperthermophilic archaeon Thermoproteus tenax a gene (treS/P) encoding a protein with similarity to annotated trehalose phosphorylase (TreP), trehalose synthase (TreS) and more recently characterized trehalose glycosyltransferring synthase (TreT) was identified. The treS/P gene as well as an upstream located ORF of unknown function (orfY) were cloned, heterologously expressed in E. coli and purified. The enzymatic characterization of the putative TreS/P revealed TreT activity. However, contrary to the previously characterized reversible TreT from Thermococcus litoralis and Pyrococcus horikoshii, the T. tenax enzyme is unidirectional and catalyzes only the formation of trehalose from UDP (ADP)-glucose and glucose. The T. tenax enzyme differs from the reversible TreT of T. litoralis by its preference for UDP-glucose as co-substrate. Phylogenetic and comparative gene context analyses reveal a conserved organization of the unidirectional TreT and OrfY gene cluster that is present in many Archaea and a few Bacteria. In contrast, the reversible TreT pathway seems to be restricted to only a few archaeal (e.g. Thermococcales) and bacterial (Thermotogales) members. Here we present a new pathway exclusively involved in trehalose synthesis––the unidirectional TreT pathway––and discuss its physiological role as well as its phylogenetic distribution.

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Abbreviations

TPS/TPP:

Trehalose-6-phosphate synthase/trehalose-6-phosphate phosphatase

TreS:

Trehalose synthase

TreT:

Trehalose glycosyltransferring synthase

TreP:

Trehalose phosphorylase

TreY/TreZ:

Maltooligosyl-trehalose synthase/maltooligosyl-trehalose trehalohydrolase

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Acknowledgments

This work was supported by the Deutsche Forschungsgemeinschaft (DFG) through the grant Si642/6-1 (SPP1112). The authors thank W. Boos (University of Konstanz, Germany) for providing the expression vector for the TreT of T. litoralis, S.V. Albers (University of Groningen, The Netherlands) for providing vector pET302 and A. Lupas (MPI for Developmental Biology, Tübingen, Germany) for support in structural analyses of OrfY.

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Authors and Affiliations

  1. Department of Chemistry, Biofilm Centre, Molecular Enzyme Technology and Biochemistry, University of Duisburg-Essen, Lotharstr., MF Building, 47057, Duisburg, Germany

    Theresa Kouril, Melanie Zaparty, Jeannette Marrero & Bettina Siebers

  2. Département de Biochimie, Faculté de Médecine, Université de Montréal, C.P. 6128, Succursale Centre-Ville, Montreal, QC, H3C3J7, Canada

    Henner Brinkmann

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  1. Theresa Kouril
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  2. Melanie Zaparty
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  3. Jeannette Marrero
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Correspondence to Bettina Siebers.

Additional information

Communicated by Harald Huber.

T. Kouril and M. Zaparty contributed equally to this work.

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Kouril, T., Zaparty, M., Marrero, J. et al. A novel trehalose synthesizing pathway in the hyperthermophilic Crenarchaeon Thermoproteus tenax: the unidirectional TreT pathway. Arch Microbiol 190, 355–369 (2008). https://doi.org/10.1007/s00203-008-0377-3

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