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AoS28D, a proline-Xaa carboxypeptidase secreted by Aspergillus oryzae

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Abstract

Prolyl peptidases of the MEROPS S28 family are of particular interest because they are key enzymes in the digestion of proline-rich peptides. A BLAST analysis of the Aspergillus oryzae genome revealed sequences coding for four proteases of the S28 family. Three of these proteases, AoS28A, AoS28B, and AoS28C, were previously characterized as acidic prolyl endopeptidases. The fourth protease, AoS28D, showed high sequence divergence with other S28 proteases and belongs to a phylogenetically distinct cluster together with orthologous proteases from other Aspergillus species. The objective of the present paper was to characterize AoS28D protease in terms of substrate specificity and activity. AoS28D produced by gene overexpression in A. oryzae and in Pichia pastoris was a 70-kDa glycoprotein with a 10-kDa sugar moiety. In contrast with other S28 proteases, AoS28D did not hydrolyze internal Pro-Xaa bonds of several tested peptides. Similarly, to human lysosomal Pro-Xaa carboxypeptidase, AoS28D demonstrated selectivity for cleaving C-terminal Pro-Xaa bonds which are resistant to carboxypeptidases of the S10 family concomitantly secreted by A. oryzae. Therefore, AoS28D could act in synergy with these enzymes during sequential degradation of a peptide from its C-terminus.

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Authors and Affiliations

  1. Service de Dermatologie, Laboratoire de Mycologie, BT422, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland

    Karine Salamin & Michel Monod

  2. Laboratoire des Catecholamines et Peptides, Service de Biomédecine, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland

    Philippe J. Eugster & Eric Grouzmann

  3. Centre for Integrative Biology, University of Trento, Trento, Italy

    Olivier Jousson

  4. Protein Analysis Facility, Center for Integrative Genomics, University of Lausanne, Lausanne, Switzerland

    Patrice Waridel

Authors
  1. Karine Salamin
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  2. Philippe J. Eugster
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  3. Olivier Jousson
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  4. Patrice Waridel
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  5. Eric Grouzmann
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  6. Michel Monod
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Correspondence to Michel Monod.

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Karine Salamin and Philippe J. Eugster contributed equally to this work.

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Salamin, K., Eugster, P.J., Jousson, O. et al. AoS28D, a proline-Xaa carboxypeptidase secreted by Aspergillus oryzae . Appl Microbiol Biotechnol 101, 4129–4137 (2017). https://doi.org/10.1007/s00253-017-8186-8

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  • DOI: https://doi.org/10.1007/s00253-017-8186-8

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