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Novel hydrolase diversity retrieved from a metagenome library of bovine rumen microflora

Authors

  • Manuel Ferrer,

    Corresponding author
    1. Department of Microbiology, GBF– German Research Centre for Biotechnology, Mascheroder Weg 1, 38124 Braunschweig, Germany.
    2. Institute of Catalysis, CSIC, Marie Curie 2, Cantoblanco, 28049 Madrid, Spain.
      E-mail mferrer@icp.csic.es; Tel. (+34) 91 585 4814; Fax (+34) 91 585 4760.
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  • Olga V. Golyshina,

    1. Department of Microbiology, GBF– German Research Centre for Biotechnology, Mascheroder Weg 1, 38124 Braunschweig, Germany.
    2. Institute of Microbiology, Technical University Braunschweig, Spielmannstr. 7, 38106 Braunschweig, Germany.
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  • Tatyana N. Chernikova,

    1. Department of Microbiology, GBF– German Research Centre for Biotechnology, Mascheroder Weg 1, 38124 Braunschweig, Germany.
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  • Amit N. Khachane,

    1. Department of Microbiology, GBF– German Research Centre for Biotechnology, Mascheroder Weg 1, 38124 Braunschweig, Germany.
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  • Dolores Reyes-Duarte,

    1. Institute of Catalysis, CSIC, Marie Curie 2, Cantoblanco, 28049 Madrid, Spain.
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  • Vitor A. P. Martins Dos Santos,

    1. Department of Microbiology, GBF– German Research Centre for Biotechnology, Mascheroder Weg 1, 38124 Braunschweig, Germany.
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  • Carsten Strompl,

    1. Department of Microbiology, GBF– German Research Centre for Biotechnology, Mascheroder Weg 1, 38124 Braunschweig, Germany.
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  • Kieran Elborough,

    1. ViaLactia Biosciences Limited, PO Box 109 185, Newmarket, Auckland, New Zealand.
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  • Graeme Jarvis,

    1. ViaLactia Biosciences Limited, PO Box 109 185, Newmarket, Auckland, New Zealand.
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      Meat and Wool New Zealand Ltd, PO Box 121, Wellington, New Zealand
  • Alexander Neef,

    1. Department of Microbiology, GBF– German Research Centre for Biotechnology, Mascheroder Weg 1, 38124 Braunschweig, Germany.
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      ‡Cavanilles Institute for Biodiversity and Evolutionary Biology, University of Valencia, Campus de Paterna, 46071 Valencia, Spain.
  • Michail M. Yakimov,

    1. Istituto per l’Ambiente Marino Costiero, CNR, Messina, Italy.
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  • Kenneth N. Timmis,

    1. Department of Microbiology, GBF– German Research Centre for Biotechnology, Mascheroder Weg 1, 38124 Braunschweig, Germany.
    2. Institute of Microbiology, Technical University Braunschweig, Spielmannstr. 7, 38106 Braunschweig, Germany.
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  • Peter N. Golyshin

    1. Department of Microbiology, GBF– German Research Centre for Biotechnology, Mascheroder Weg 1, 38124 Braunschweig, Germany.
    2. Institute of Microbiology, Technical University Braunschweig, Spielmannstr. 7, 38106 Braunschweig, Germany.
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  • Present Addresses: Meat and Wool New Zealand Ltd, PO Box 121, Wellington, New Zealand; Cavanilles Institute for Biodiversity and Evolutionary Biology, University of Valencia, Campus de Paterna, 46071 Valencia, Spain.

E-mail mferrer@icp.csic.es; Tel. (+34) 91 585 4814; Fax (+34) 91 585 4760.

Summary

A metagenome expression library of bulk DNA extracted from the rumen content of a dairy cow was established in a phage lambda vector and activity-based screening employed to explore the functional diversity of the microbial flora. Twenty-two clones specifying distinct hydrolytic activities (12 esterases, nine endo-β-1,4-glucanases and one cyclodextrinase) were identified in the library and characterized. Sequence analysis of the retrieved enzymes revealed that eight (36%) were entirely new and formed deep-branched phylogenetic lineages with no close relatives among known ester- and glycosyl-hydrolases. Bioinformatic analyses of the hydrolase gene sequences, and the sequences and contexts of neighbouring genes, suggested tentative phylogenetic assignments of the rumen organisms producing the retrieved enzymes. The phylogenetic novelty of the hydrolases suggests that some of them may have potential for new applications in biocatalysis.

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