Cloning and sequencing of two Ceriporiopsis subvermispora bicupin oxalate oxidase allelic isoforms: Implications for the reaction specificity of oxalate oxidases and decarboxylases

Escutia, Marta R., Bowater, Laura, Edwards, Anne, Bottrill, Andrew R., Burrell, Matthew R., Polanco, Rubén, Vicuña, Rafael and Bornemann, Stephen (2005) Cloning and sequencing of two Ceriporiopsis subvermispora bicupin oxalate oxidase allelic isoforms: Implications for the reaction specificity of oxalate oxidases and decarboxylases. Applied and Environmental Microbiology, 71 (7). pp. 3608-3616. ISSN 0099-2240

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Abstract

Oxalate oxidase is thought to be involved in the production of hydrogen peroxide for lignin degradation by the dikaryotic white rot fungus Ceriporiopsis subvermispora. This enzyme was purified, and after digestion with trypsin, peptide fragments of the enzyme were sequenced using quadrupole time-of-flight mass spectrometry. Starting with degenerate primers based on the peptide sequences, two genes encoding isoforms of the enzyme were cloned, sequenced, and shown to be allelic. Both genes contained 14 introns. The sequences of the isoforms revealed that they were both bicupins that unexpectedly shared the greatest similarity to microbial bicupin oxalate decarboxylases rather than monocupin plant oxalate oxidases (also known as germins). We have shown that both fungal isoforms, one of which was heterologously expressed in Escherichia coli, are indeed oxalate oxidases that possess ≤0.2% oxalate decarboxylase activity and that the organism is capable of rapidly degrading exogenously supplied oxalate. They are therefore the first bicupin oxalate oxidases to have been described. Heterologous expression of active enzyme was dependent on the addition of manganese salts to the growth medium. Molecular modeling provides new and independent evidence for the identity of the catalytic site and the key amino acid involved in defining the reaction specificities of oxalate oxidases and oxalate decarboxylases.

Item Type:	Article
Faculty \ School:	Faculty of Medicine and Health Sciences > Norwich Medical School Faculty of Science > School of Biological Sciences Faculty of Science > School of Chemical Sciences and Pharmacy (former - to 2009)
Depositing User:	Rhiannon Harvey
Date Deposited:	31 May 2011 11:21
Last Modified:	24 Jan 2024 01:22
URI:	https://ueaeprints.uea.ac.uk/id/eprint/31573
DOI:	10.1128/AEM.71.7.3608-3616.2005

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