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Purification and characterization of an extracellular laccase from solid-state culture of Pleurotus ostreatus HP-1
A native isolate of Pleurotus ostreatus HP-1 (Genbank Accession No. EU420068) was found to have an excellent laccase producing ability. The extracellular laccase was purified to electrophoretic homogeneity from copper sulphate induced solid-state fermentation medium by ammonium sulphate precipitatio...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3909575/ https://www.ncbi.nlm.nih.gov/pubmed/28324461 http://dx.doi.org/10.1007/s13205-013-0129-1 |
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author | Patel, Hardik Gupte, Shilpa Gahlout, Mayur Gupte, Akshaya |
author_facet | Patel, Hardik Gupte, Shilpa Gahlout, Mayur Gupte, Akshaya |
author_sort | Patel, Hardik |
collection | PubMed |
description | A native isolate of Pleurotus ostreatus HP-1 (Genbank Accession No. EU420068) was found to have an excellent laccase producing ability. The extracellular laccase was purified to electrophoretic homogeneity from copper sulphate induced solid-state fermentation medium by ammonium sulphate precipitation and ion-exchange chromatography. The enzyme was determined to be monomeric protein with an apparent molecular mass of 68,420 kDa, and an isoelectric point (pI) of 3.5. The inductively coupled plasma spectroscopy showed a presence of iron, zinc and copper in the purified enzyme. The absorption spectrum in the range of 200–700 nm showed the maximum absorption at 610 nm characteristic of fungal laccase and corresponding to the presence of type I copper atom. The laccase was stable at different temperatures up to 70 °C and retained 61 % activity at 50 °C. The enzyme reaction was inhibited by cysteine; sodium azide and EDTA. The enzyme oxidized various known laccase substrates, its lowest K(m) value being for ortho-dianisidine and highest K(cat) and K(cat)/K(m) for ABTS. The purified laccase exhibited different pH optima for different substrates. The N-terminal sequence did not show any similarity with N-terminal sequence of other species of genera Pleurotus. |
format | Online Article Text |
id | pubmed-3909575 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-39095752014-02-06 Purification and characterization of an extracellular laccase from solid-state culture of Pleurotus ostreatus HP-1 Patel, Hardik Gupte, Shilpa Gahlout, Mayur Gupte, Akshaya 3 Biotech Original Article A native isolate of Pleurotus ostreatus HP-1 (Genbank Accession No. EU420068) was found to have an excellent laccase producing ability. The extracellular laccase was purified to electrophoretic homogeneity from copper sulphate induced solid-state fermentation medium by ammonium sulphate precipitation and ion-exchange chromatography. The enzyme was determined to be monomeric protein with an apparent molecular mass of 68,420 kDa, and an isoelectric point (pI) of 3.5. The inductively coupled plasma spectroscopy showed a presence of iron, zinc and copper in the purified enzyme. The absorption spectrum in the range of 200–700 nm showed the maximum absorption at 610 nm characteristic of fungal laccase and corresponding to the presence of type I copper atom. The laccase was stable at different temperatures up to 70 °C and retained 61 % activity at 50 °C. The enzyme reaction was inhibited by cysteine; sodium azide and EDTA. The enzyme oxidized various known laccase substrates, its lowest K(m) value being for ortho-dianisidine and highest K(cat) and K(cat)/K(m) for ABTS. The purified laccase exhibited different pH optima for different substrates. The N-terminal sequence did not show any similarity with N-terminal sequence of other species of genera Pleurotus. Springer Berlin Heidelberg 2013-03-16 2014-02 /pmc/articles/PMC3909575/ /pubmed/28324461 http://dx.doi.org/10.1007/s13205-013-0129-1 Text en © The Author(s) 2013 https://creativecommons.org/licenses/by/2.0/ This article is published under license to BioMed Central Ltd.Open AccessThis article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited. |
spellingShingle | Original Article Patel, Hardik Gupte, Shilpa Gahlout, Mayur Gupte, Akshaya Purification and characterization of an extracellular laccase from solid-state culture of Pleurotus ostreatus HP-1 |
title | Purification and characterization of an extracellular laccase from solid-state culture of Pleurotus ostreatus HP-1 |
title_full | Purification and characterization of an extracellular laccase from solid-state culture of Pleurotus ostreatus HP-1 |
title_fullStr | Purification and characterization of an extracellular laccase from solid-state culture of Pleurotus ostreatus HP-1 |
title_full_unstemmed | Purification and characterization of an extracellular laccase from solid-state culture of Pleurotus ostreatus HP-1 |
title_short | Purification and characterization of an extracellular laccase from solid-state culture of Pleurotus ostreatus HP-1 |
title_sort | purification and characterization of an extracellular laccase from solid-state culture of pleurotus ostreatus hp-1 |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3909575/ https://www.ncbi.nlm.nih.gov/pubmed/28324461 http://dx.doi.org/10.1007/s13205-013-0129-1 |
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