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The crystal structure of an extracellular catechol oxidase from the ascomycete fungus Aspergillus oryzae

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Abstract

Catechol oxidases (EC 1.10.3.1) catalyse the oxidation of o-diphenols to their corresponding o-quinones. These oxidases contain two copper ions (CuA and CuB) within the so-called coupled type 3 copper site as found in tyrosinases (EC 1.14.18.1) and haemocyanins. The crystal structures of a limited number of bacterial and fungal tyrosinases and plant catechol oxidases have been solved. In this study, we present the first crystal structure of a fungal catechol oxidase from Aspergillus oryzae (AoCO4) at 2.5-Å resolution. AoCO4 belongs to the newly discovered family of short-tyrosinases, which are distinct from other tyrosinases and catechol oxidases because of their lack of the conserved C-terminal domain and differences in the histidine pattern for CuA. The sequence identity of AoCO4 with other structurally known enzymes is low (less than 30 %), and the crystal structure of AoCO4 diverges from that of enzymes belonging to the conventional tyrosinase family in several ways, particularly around the central α-helical core region. A diatomic oxygen moiety was identified as a bridging molecule between the two copper ions CuA and CuB separated by a distance of 4.2–4.3 Å. The UV/vis absorption spectrum of AoCO4 exhibits a distinct maximum of absorbance at 350 nm, which has been reported to be typical of the oxy form of type 3 copper enzymes.

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Abbreviations

AbT:

Tyrosinase from Agaricus bisporus

AoCO4:

Catechol oxidase from the fungus Aspergillus oryzae

BmT:

Tyrosinase from Bacillus megaterium

DESY:

Deutsches Elektronen-Synchrotron

ESRF:

European Synchrotron Radiation Facility

IbCO:

Catechol oxidase from Ipomea batatas

NAG:

N-Acetylglucosamine

ScT:

Tyrosinase from Streptomyces castaneoglobisporus

T3:

Type 3

VvCO:

Catechol oxidase from Vitis vinifera

MAN:

Mannose

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Acknowledgments

The work was conducted with financial support from the Academy of Finland (projects 115085, 256937 and 263931), the Marie Curie EU project “Enzymatic tailoring of protein interactions and functionalities in food matrix” PRO-ENZ (MEST-CT-2005-020924) and the Research Foundation of Raisiogroup (Raisio, Finland). The ESRF, Grenoble, and DESY, Hamburg, are thanked for their provision of the synchrotron facilities.

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Authors and Affiliations

  1. Department of Chemistry, University of Eastern Finland, Joensuu Campus, PO Box 111, 80101, Joensuu, Finland

    Nina Hakulinen, Chiara Gasparetti, Heidi Kaljunen & Juha Rouvinen

  2. VTT Technical Research Center of Finland, PO Box 1000, 02044, Espoo, Finland

    Chiara Gasparetti & Kristiina Kruus

  3. EMBL Hamburg c/o DESY, Notkestraße 85, 22603, Hamburg, Germany

    Heidi Kaljunen

Authors
  1. Nina Hakulinen
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  2. Chiara Gasparetti
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  3. Heidi Kaljunen
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  4. Kristiina Kruus
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  5. Juha Rouvinen
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Corresponding author

Correspondence to Nina Hakulinen.

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An interactive 3D complement page in Proteopedia is available at http://proteopedia.org/w/Journal:JBIC:22.

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Hakulinen, N., Gasparetti, C., Kaljunen, H. et al. The crystal structure of an extracellular catechol oxidase from the ascomycete fungus Aspergillus oryzae . J Biol Inorg Chem 18, 917–929 (2013). https://doi.org/10.1007/s00775-013-1038-9

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  • DOI: https://doi.org/10.1007/s00775-013-1038-9

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