Identification and characterization of a novel multicopper oxidase from Acidomyces acidophilus with ferroxidase activity

Research output: Contribution to journalArticle

Abstract

A new multicopper oxidase gene AaMco1 was identified in Acidomyces acidophilus, a pigmented extremophile ascomycete originally isolated from acidic water. Sequence analysis revealed that it encodes a 682 amino acid protein with an apparent molecular mass of 85 kDa as determined by denaturing SDS-PAGE. Interestingly, AaMco1 has a predicted N-terminal transmembrane helix and no signal peptide. To obtain an active and soluble protein, AaMco1 was truncated at its N-terminal to remove the transmembrane helix, but even in this form the protein was found in the insoluble fraction. AaMco1 and its truncated form were then denatured, purified and renatured before characterization. Structural analysis and protein characterization by enzymatic assays indicate that AaMco1 has ferroxidase activity. AaMco1 is also able to oxidize the DMPPDA compound and could be part of a new phylogenetic cluster, the ascomycete MCOs family, described for the first time here.

Original languageEnglish
Pages (from-to)37-46
JournalBiochimie
Volume102
DOIs
Publication statusPublished - 13 Mar 2014

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Ceruloplasmin
Oxidoreductases
Ascomycota
Proteins
Enzyme Assays
Molecular mass
Protein Sorting Signals
Structural analysis
Sequence Analysis
Polyacrylamide Gel Electrophoresis
Assays
Genes
Amino Acids
Water

Keywords

  • Acidomyces acidophilus
  • Ferroxidase activity
  • N-terminal transmembrane helix
  • New multicopper oxidase

Cite this

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title = "Identification and characterization of a novel multicopper oxidase from Acidomyces acidophilus with ferroxidase activity",
abstract = "A new multicopper oxidase gene AaMco1 was identified in Acidomyces acidophilus, a pigmented extremophile ascomycete originally isolated from acidic water. Sequence analysis revealed that it encodes a 682 amino acid protein with an apparent molecular mass of 85 kDa as determined by denaturing SDS-PAGE. Interestingly, AaMco1 has a predicted N-terminal transmembrane helix and no signal peptide. To obtain an active and soluble protein, AaMco1 was truncated at its N-terminal to remove the transmembrane helix, but even in this form the protein was found in the insoluble fraction. AaMco1 and its truncated form were then denatured, purified and renatured before characterization. Structural analysis and protein characterization by enzymatic assays indicate that AaMco1 has ferroxidase activity. AaMco1 is also able to oxidize the DMPPDA compound and could be part of a new phylogenetic cluster, the ascomycete MCOs family, described for the first time here.",
keywords = "Acidomyces acidophilus, Ferroxidase activity, N-terminal transmembrane helix, New multicopper oxidase",
author = "France Boonen and Anne-Mich{\`e}le Vandamme and Emilie Etoundi and Lise-Marie Pigneur and Isabelle Housen",
year = "2014",
month = "3",
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language = "English",
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journal = "Biochimie",
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TY - JOUR

T1 - Identification and characterization of a novel multicopper oxidase from Acidomyces acidophilus with ferroxidase activity

AU - Boonen, France

AU - Vandamme, Anne-Michèle

AU - Etoundi, Emilie

AU - Pigneur, Lise-Marie

AU - Housen, Isabelle

PY - 2014/3/13

Y1 - 2014/3/13

N2 - A new multicopper oxidase gene AaMco1 was identified in Acidomyces acidophilus, a pigmented extremophile ascomycete originally isolated from acidic water. Sequence analysis revealed that it encodes a 682 amino acid protein with an apparent molecular mass of 85 kDa as determined by denaturing SDS-PAGE. Interestingly, AaMco1 has a predicted N-terminal transmembrane helix and no signal peptide. To obtain an active and soluble protein, AaMco1 was truncated at its N-terminal to remove the transmembrane helix, but even in this form the protein was found in the insoluble fraction. AaMco1 and its truncated form were then denatured, purified and renatured before characterization. Structural analysis and protein characterization by enzymatic assays indicate that AaMco1 has ferroxidase activity. AaMco1 is also able to oxidize the DMPPDA compound and could be part of a new phylogenetic cluster, the ascomycete MCOs family, described for the first time here.

AB - A new multicopper oxidase gene AaMco1 was identified in Acidomyces acidophilus, a pigmented extremophile ascomycete originally isolated from acidic water. Sequence analysis revealed that it encodes a 682 amino acid protein with an apparent molecular mass of 85 kDa as determined by denaturing SDS-PAGE. Interestingly, AaMco1 has a predicted N-terminal transmembrane helix and no signal peptide. To obtain an active and soluble protein, AaMco1 was truncated at its N-terminal to remove the transmembrane helix, but even in this form the protein was found in the insoluble fraction. AaMco1 and its truncated form were then denatured, purified and renatured before characterization. Structural analysis and protein characterization by enzymatic assays indicate that AaMco1 has ferroxidase activity. AaMco1 is also able to oxidize the DMPPDA compound and could be part of a new phylogenetic cluster, the ascomycete MCOs family, described for the first time here.

KW - Acidomyces acidophilus

KW - Ferroxidase activity

KW - N-terminal transmembrane helix

KW - New multicopper oxidase

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DO - 10.1016/j.biochi.2014.02.009

M3 - Article

VL - 102

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JO - Biochimie

JF - Biochimie

SN - 0300-9084

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