Purification, characterization and sequence analysis of a laccase from the ascomycete Mauginiella sp.

Hetti Palonen, Markku Saloheimo, Liisa Viikari, Kristiina Kruus (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

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Abstract

A laccase from the ascomycete Mauginiella sp. was purified to electrophoretic homogeneity and biochemically characterized. The molecular mass of the laccase was 63 kDa as determined by mass spectrometry and it existed as six isoforms with isoelectric points in the range of 4.8–6.4. The laccase showed activity towards the typical substrates: 2,2′-azinobis-(3-ethylbenzothiazoline)-6-sulphonate (ABTS), guaiacol, dimethoxyphenol (2,6-DMP), and syringaldazine. The pH optima on guaiacol, 2,6-DMP and ABTS were 4, 3.5 and 2.4, respectively. The enzyme was strongly, 98%, inhibited by 1 mM NaN3 and 88% by 1 mM KCN. The laccase was stable at neutral pH, retaining 80% activity after 24 h at pH 6–8. The enzyme was sensitive to high temperatures: the half-life at 70 °C was only 3 min. A fragment of the laccase gene was isolated and its nucleotide sequence was determined. The laccase gene showed high identity to the laccase genes lcc1 and lcc2 of the basidiomycetes Trametes versicolor and Trametes villosa, respectively.
Original languageEnglish
Pages (from-to)854-862
Number of pages9
JournalEnzyme and Microbial Technology
Volume33
Issue number6
DOIs
Publication statusPublished - 2003
MoE publication typeA1 Journal article-refereed

Fingerprint

Laccase
Ascomycota
Purification
Sequence Analysis
Genes
Enzymes
Trametes
Guaiacol
Molecular mass
Nucleotides
Mass spectrometry
Basidiomycota
Sodium Azide
Substrates
Isoelectric Point
Half-Life
Mass Spectrometry
Protein Isoforms
Temperature

Keywords

  • Laccase
  • Mauginiella sp.
  • Purification
  • Characterization
  • Amino acid sequence
  • Gene sequence

Cite this

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title = "Purification, characterization and sequence analysis of a laccase from the ascomycete Mauginiella sp.",
abstract = "A laccase from the ascomycete Mauginiella sp. was purified to electrophoretic homogeneity and biochemically characterized. The molecular mass of the laccase was 63 kDa as determined by mass spectrometry and it existed as six isoforms with isoelectric points in the range of 4.8–6.4. The laccase showed activity towards the typical substrates: 2,2′-azinobis-(3-ethylbenzothiazoline)-6-sulphonate (ABTS), guaiacol, dimethoxyphenol (2,6-DMP), and syringaldazine. The pH optima on guaiacol, 2,6-DMP and ABTS were 4, 3.5 and 2.4, respectively. The enzyme was strongly, 98{\%}, inhibited by 1 mM NaN3 and 88{\%} by 1 mM KCN. The laccase was stable at neutral pH, retaining 80{\%} activity after 24 h at pH 6–8. The enzyme was sensitive to high temperatures: the half-life at 70 °C was only 3 min. A fragment of the laccase gene was isolated and its nucleotide sequence was determined. The laccase gene showed high identity to the laccase genes lcc1 and lcc2 of the basidiomycetes Trametes versicolor and Trametes villosa, respectively.",
keywords = "Laccase, Mauginiella sp., Purification, Characterization, Amino acid sequence, Gene sequence",
author = "Hetti Palonen and Markku Saloheimo and Liisa Viikari and Kristiina Kruus",
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Purification, characterization and sequence analysis of a laccase from the ascomycete Mauginiella sp. / Palonen, Hetti; Saloheimo, Markku; Viikari, Liisa; Kruus, Kristiina (Corresponding Author).

In: Enzyme and Microbial Technology, Vol. 33, No. 6, 2003, p. 854-862.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Purification, characterization and sequence analysis of a laccase from the ascomycete Mauginiella sp.

AU - Palonen, Hetti

AU - Saloheimo, Markku

AU - Viikari, Liisa

AU - Kruus, Kristiina

PY - 2003

Y1 - 2003

N2 - A laccase from the ascomycete Mauginiella sp. was purified to electrophoretic homogeneity and biochemically characterized. The molecular mass of the laccase was 63 kDa as determined by mass spectrometry and it existed as six isoforms with isoelectric points in the range of 4.8–6.4. The laccase showed activity towards the typical substrates: 2,2′-azinobis-(3-ethylbenzothiazoline)-6-sulphonate (ABTS), guaiacol, dimethoxyphenol (2,6-DMP), and syringaldazine. The pH optima on guaiacol, 2,6-DMP and ABTS were 4, 3.5 and 2.4, respectively. The enzyme was strongly, 98%, inhibited by 1 mM NaN3 and 88% by 1 mM KCN. The laccase was stable at neutral pH, retaining 80% activity after 24 h at pH 6–8. The enzyme was sensitive to high temperatures: the half-life at 70 °C was only 3 min. A fragment of the laccase gene was isolated and its nucleotide sequence was determined. The laccase gene showed high identity to the laccase genes lcc1 and lcc2 of the basidiomycetes Trametes versicolor and Trametes villosa, respectively.

AB - A laccase from the ascomycete Mauginiella sp. was purified to electrophoretic homogeneity and biochemically characterized. The molecular mass of the laccase was 63 kDa as determined by mass spectrometry and it existed as six isoforms with isoelectric points in the range of 4.8–6.4. The laccase showed activity towards the typical substrates: 2,2′-azinobis-(3-ethylbenzothiazoline)-6-sulphonate (ABTS), guaiacol, dimethoxyphenol (2,6-DMP), and syringaldazine. The pH optima on guaiacol, 2,6-DMP and ABTS were 4, 3.5 and 2.4, respectively. The enzyme was strongly, 98%, inhibited by 1 mM NaN3 and 88% by 1 mM KCN. The laccase was stable at neutral pH, retaining 80% activity after 24 h at pH 6–8. The enzyme was sensitive to high temperatures: the half-life at 70 °C was only 3 min. A fragment of the laccase gene was isolated and its nucleotide sequence was determined. The laccase gene showed high identity to the laccase genes lcc1 and lcc2 of the basidiomycetes Trametes versicolor and Trametes villosa, respectively.

KW - Laccase

KW - Mauginiella sp.

KW - Purification

KW - Characterization

KW - Amino acid sequence

KW - Gene sequence

U2 - 10.1016/S0141-0229(03)00247-3

DO - 10.1016/S0141-0229(03)00247-3

M3 - Article

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JO - Enzyme and Microbial Technology

JF - Enzyme and Microbial Technology

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