Structure, recombinant expression and mutagenesis studies of the catalase with oxidase activity from Scytalidium thermophilum

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Date

2013-03-01

Author

YÜZÜGÜLLÜ, YONCA
Trinh, Chi H.
Smith, Mark A.
Pearson, Arwen R.
Phillips, Simon E. V.
SUTAY KOCABAŞ, DİDEM
Bakir, Ufuk
Ögel, Zümrüt Begüm
McPherson, Michael J.

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Scytalidium thermophilum produces a catalase with phenol oxidase activity (CATPO) that catalyses the decomposition of hydrogen peroxide into oxygen and water and also oxidizes various phenolic compounds. A codon-optimized catpo gene was cloned and expressed in Escherichia coli. The crystal structures of native and recombinant S. thermophilum CATPO and two variants, H82N and V123F, were determined at resolutions of 2.7, 1.4, 1.5 and 1.9 angstrom, respectively. The structure of CATPO reveals a homotetramer with 698 residues per subunit and with strong structural similarity to Penicillium vitale catalase. The haem component is cis-hydroxychlorin -spirolactone, which is rotated 180 degrees with respect to small-subunit catalases. The haem-binding pocket contains two highly conserved water molecules on the distal side. The H82N mutation resulted in conversion of the native d-type haem to a b-type haem. Kinetic studies of the H82N and V123F mutants indicate that both activities are likely to be associated with the haem centre and suggest that the secondary oxidase activity may be a general feature of catalases in the absence of hydrogen peroxide.

Subject Keywords

Structural Biology, General Medicine

URI

https://hdl.handle.net/11511/56517

Journal

ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY

DOI

https://doi.org/10.1107/s0907444912049001

Collections

Graduate School of Natural and Applied Sciences, Article

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Citation Formats

Y. YÜZÜGÜLLÜ et al., “Structure, recombinant expression and mutagenesis studies of the catalase with oxidase activity from Scytalidium thermophilum,” ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, pp. 398–408, 2013, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/56517.