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Applied Microbiology and Biotechnology

, Volume 90, Issue 3, pp 941–949 | Cite as

Production and characterisation of AoSOX2 from Aspergillus oryzae, a novel flavin-dependent sulfhydryl oxidase with good pH and temperature stability

  • Greta Faccio
  • Kristiina Kruus
  • Johanna Buchert
  • Markku Saloheimo
Biotechnologically Relevant Enzymes and Proteins

Abstract

Sulfhydryl oxidases have found application in the improvement of both dairy and baking products due to their ability to oxidise thiol groups in small molecules and cysteine residues in proteins. A genome mining study of the available fungal genomes had previously been performed by our group in order to identify novel sulfhydryl oxidases suitable for industrial applications and a representative enzyme was produced, AoSOX1 from Aspergillus oryzae (Faccio et al. BMC Biochem 11:31, 2010). As a result of the study, a second gene coding for a potentially secreted sulfhydryl oxidase, AoSOX2, was identified in the genome of A. oryzae. The protein AoSOX2 was heterologously expressed in Trichoderma reesei and characterised with regard to both biochemical properties as well as preliminary structural analysis. AoSOX2 showed activity on dithiothreitol and glutathione, and to a lesser extent on D/L-cysteine and beta-mercaptoethanol. AoSOX2 was a homodimeric flavin-dependent protein of approximately 78 kDa (monomer 42412 Da) and its secondary structure presents alpha-helical elements. A. oryzae AoSOX2 showed a significant stability to pH and temperature.

Keywords

Sulfhydryl oxidase Secreted Fungal Flavoenzyme Production Characterization 

Notes

Acknowledgements

The work was funded by the Marie Curie EU-project PRO-ENZ (MEST-CT-2005-020924) and by a personal grant to GF by the Finnish Cultural Foundation. We are thankful to Harry Boer and Evanthia Monogioudi for the assistance in CD spectroscopy and mass spectrometry. The technical assistance of Hanna Kuusinen is also acknowledged. The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Greta Faccio
    • 1
  • Kristiina Kruus
    • 1
  • Johanna Buchert
    • 1
  • Markku Saloheimo
    • 1
  1. 1.VTT Technical Research Centre of FinlandEspooFinland

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