Purification and characterisation of a novel laccase from the ascomycete Melanocarpus albomyces

Laura-Leena Kiiskinen, Liisa Viikari, Kristiina Kruus

Research output: Contribution to journalArticleScientificpeer-review

176 Citations (Scopus)

Abstract

A novel laccase from the ascomycete Melanocarpus albomyces was purified and characterised. The enzyme was purified using anion exchange chromatography, hydrophobic interaction chromatography and gel filtration, and the purified laccase was biochemically characterised. It had activity towards typical substrates of laccases including 2,2-azinobis-(3-ethylbenzthiazoline-6-sulphonate), dimethoxyphenol, guaiacol, and syringaldazine. The laccase showed good thermostability and it had a pH optimum at neutral pH, both unusual properties for most known fungal laccases. The activity of the laccase from M. albomyces was highest at 60-70°C. With guaiacol and syringaldazine the pH optima were rather broad: 5-7.5 and 6-7, respectively. It retained 50% of its activity after 5 h incubation at 60°C. The molecular weight of the laccase was about 80 kDa and the isoelectric point 4.0. The ultraviolet-visible absorption and electron paramagnetic resonance spectra of the purified laccase indicated that the typical three types of copper were present.
Original languageEnglish
Pages (from-to)198-204
JournalApplied Microbiology and Biotechnology
Volume59
Issue number2-3
DOIs
Publication statusPublished - 2002
MoE publication typeA1 Journal article-refereed

Fingerprint

Laccase
Ascomycota
Guaiacol
Electron Spin Resonance Spectroscopy
Isoelectric Point
Hydrophobic and Hydrophilic Interactions
Gel Chromatography
Anions
Chromatography
Copper
Molecular Weight
Enzymes

Keywords

  • laccase
  • ascomycete
  • Melanocarpus albomyces
  • anion exchange chromatography
  • hydrophobic interaction chromatography
  • gel filtration
  • enzymes
  • oxidases

Cite this

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title = "Purification and characterisation of a novel laccase from the ascomycete Melanocarpus albomyces",
abstract = "A novel laccase from the ascomycete Melanocarpus albomyces was purified and characterised. The enzyme was purified using anion exchange chromatography, hydrophobic interaction chromatography and gel filtration, and the purified laccase was biochemically characterised. It had activity towards typical substrates of laccases including 2,2-azinobis-(3-ethylbenzthiazoline-6-sulphonate), dimethoxyphenol, guaiacol, and syringaldazine. The laccase showed good thermostability and it had a pH optimum at neutral pH, both unusual properties for most known fungal laccases. The activity of the laccase from M. albomyces was highest at 60-70°C. With guaiacol and syringaldazine the pH optima were rather broad: 5-7.5 and 6-7, respectively. It retained 50{\%} of its activity after 5 h incubation at 60°C. The molecular weight of the laccase was about 80 kDa and the isoelectric point 4.0. The ultraviolet-visible absorption and electron paramagnetic resonance spectra of the purified laccase indicated that the typical three types of copper were present.",
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Purification and characterisation of a novel laccase from the ascomycete Melanocarpus albomyces. / Kiiskinen, Laura-Leena; Viikari, Liisa; Kruus, Kristiina.

In: Applied Microbiology and Biotechnology, Vol. 59, No. 2-3, 2002, p. 198-204.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Purification and characterisation of a novel laccase from the ascomycete Melanocarpus albomyces

AU - Kiiskinen, Laura-Leena

AU - Viikari, Liisa

AU - Kruus, Kristiina

PY - 2002

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N2 - A novel laccase from the ascomycete Melanocarpus albomyces was purified and characterised. The enzyme was purified using anion exchange chromatography, hydrophobic interaction chromatography and gel filtration, and the purified laccase was biochemically characterised. It had activity towards typical substrates of laccases including 2,2-azinobis-(3-ethylbenzthiazoline-6-sulphonate), dimethoxyphenol, guaiacol, and syringaldazine. The laccase showed good thermostability and it had a pH optimum at neutral pH, both unusual properties for most known fungal laccases. The activity of the laccase from M. albomyces was highest at 60-70°C. With guaiacol and syringaldazine the pH optima were rather broad: 5-7.5 and 6-7, respectively. It retained 50% of its activity after 5 h incubation at 60°C. The molecular weight of the laccase was about 80 kDa and the isoelectric point 4.0. The ultraviolet-visible absorption and electron paramagnetic resonance spectra of the purified laccase indicated that the typical three types of copper were present.

AB - A novel laccase from the ascomycete Melanocarpus albomyces was purified and characterised. The enzyme was purified using anion exchange chromatography, hydrophobic interaction chromatography and gel filtration, and the purified laccase was biochemically characterised. It had activity towards typical substrates of laccases including 2,2-azinobis-(3-ethylbenzthiazoline-6-sulphonate), dimethoxyphenol, guaiacol, and syringaldazine. The laccase showed good thermostability and it had a pH optimum at neutral pH, both unusual properties for most known fungal laccases. The activity of the laccase from M. albomyces was highest at 60-70°C. With guaiacol and syringaldazine the pH optima were rather broad: 5-7.5 and 6-7, respectively. It retained 50% of its activity after 5 h incubation at 60°C. The molecular weight of the laccase was about 80 kDa and the isoelectric point 4.0. The ultraviolet-visible absorption and electron paramagnetic resonance spectra of the purified laccase indicated that the typical three types of copper were present.

KW - laccase

KW - ascomycete

KW - Melanocarpus albomyces

KW - anion exchange chromatography

KW - hydrophobic interaction chromatography

KW - gel filtration

KW - enzymes

KW - oxidases

U2 - 10.1007/s00253-002-1012-x

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JO - Applied Microbiology and Biotechnology

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SN - 0175-7598

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