Molecular characterization of a novel thermostable laccase PPLCC2 from the brown rot fungus Postia placenta MAD-698-R
Background: Laccase has been considered important for the degradation of lignocellulose by wood rot fungi. The properties and functions of laccase in white rot fungi have been investigated extensively, but those from brown rot fungi remain largely unknown. In this paper, a laccase isoform Pplcc2 fro...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2015-11-01
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| Series: | Electronic Journal of Biotechnology |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S0717345815001207 |
| _version_ | 1819068454172360704 |
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| author | Hongde An Tingting Xiao Huan Fan Dongsheng Wei |
| author_facet | Hongde An Tingting Xiao Huan Fan Dongsheng Wei |
| author_sort | Hongde An |
| collection | DOAJ |
| description | Background: Laccase has been considered important for the degradation of lignocellulose by wood rot fungi. The properties and functions of laccase in white rot fungi have been investigated extensively, but those from brown rot fungi remain largely unknown. In this paper, a laccase isoform Pplcc2 from the brown rot fungus Postia placenta MAD-698-R was expressed heterologously in Pichia pastoris GS115, purified and the properties of the enzyme were determined.
Results: The molecular weight of the protein was determined to be 67 kDa using SDS-PAGE. It cannot oxidize syringaldazine (SGZ), but it can oxidize 2,2′-azino-di-(3-ethylbenzothialozin-6-Sulfonic acid) (ABTS) and 2,6-dimethoxyphenol (DMP). Specific activity for ABTS was 1960 ± 19 Unit/mg. The catalytic constant (kcat) was 1213 ± 18.3 s-1 for ABTS and 293.2 ± 21.9 s-1 for DMP. Km was 22.08 μM for ABTS and 11.62 μM for DMP. The optimal pH for the oxidation of ABTS and DMP was 3.5 and 5.0 respectively. The optimal temperature for the oxidation of ABTS and DMP was 60°C.
Conclusions: This is the first identified thermo activated and thermostable laccase in brown rot fungi. This investigation will contribute to understanding the roles played by laccases in brown rot fungi. |
| first_indexed | 2024-12-21T16:34:24Z |
| format | Article |
| id | doaj.art-74af8aebc50a409694cd356c64194fc9 |
| institution | Directory Open Access Journal |
| issn | 0717-3458 |
| language | English |
| last_indexed | 2024-12-21T16:34:24Z |
| publishDate | 2015-11-01 |
| publisher | Elsevier |
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| series | Electronic Journal of Biotechnology |
| spelling | doaj.art-74af8aebc50a409694cd356c64194fc92022-12-21T18:57:15ZengElsevierElectronic Journal of Biotechnology0717-34582015-11-0118645145810.1016/j.ejbt.2015.09.004Molecular characterization of a novel thermostable laccase PPLCC2 from the brown rot fungus Postia placenta MAD-698-RHongde An0Tingting Xiao1Huan Fan2Dongsheng Wei3Department of Microbiology, College of Life Science, Nankai University, Tianjin, PR ChinaDepartment of Microbiology, College of Life Science, Nankai University, Tianjin, PR ChinaTianjin Institute of Animal Husbandry and Veterinary Research, Tianjin, PR ChinaDepartment of Microbiology, College of Life Science, Nankai University, Tianjin, PR ChinaBackground: Laccase has been considered important for the degradation of lignocellulose by wood rot fungi. The properties and functions of laccase in white rot fungi have been investigated extensively, but those from brown rot fungi remain largely unknown. In this paper, a laccase isoform Pplcc2 from the brown rot fungus Postia placenta MAD-698-R was expressed heterologously in Pichia pastoris GS115, purified and the properties of the enzyme were determined. Results: The molecular weight of the protein was determined to be 67 kDa using SDS-PAGE. It cannot oxidize syringaldazine (SGZ), but it can oxidize 2,2′-azino-di-(3-ethylbenzothialozin-6-Sulfonic acid) (ABTS) and 2,6-dimethoxyphenol (DMP). Specific activity for ABTS was 1960 ± 19 Unit/mg. The catalytic constant (kcat) was 1213 ± 18.3 s-1 for ABTS and 293.2 ± 21.9 s-1 for DMP. Km was 22.08 μM for ABTS and 11.62 μM for DMP. The optimal pH for the oxidation of ABTS and DMP was 3.5 and 5.0 respectively. The optimal temperature for the oxidation of ABTS and DMP was 60°C. Conclusions: This is the first identified thermo activated and thermostable laccase in brown rot fungi. This investigation will contribute to understanding the roles played by laccases in brown rot fungi.http://www.sciencedirect.com/science/article/pii/S0717345815001207Brown rot fungiPichia pastorisPostia placentaThermostable laccase |
| spellingShingle | Hongde An Tingting Xiao Huan Fan Dongsheng Wei Molecular characterization of a novel thermostable laccase PPLCC2 from the brown rot fungus Postia placenta MAD-698-R Electronic Journal of Biotechnology Brown rot fungi Pichia pastoris Postia placenta Thermostable laccase |
| title | Molecular characterization of a novel thermostable laccase PPLCC2 from the brown rot fungus Postia placenta MAD-698-R |
| title_full | Molecular characterization of a novel thermostable laccase PPLCC2 from the brown rot fungus Postia placenta MAD-698-R |
| title_fullStr | Molecular characterization of a novel thermostable laccase PPLCC2 from the brown rot fungus Postia placenta MAD-698-R |
| title_full_unstemmed | Molecular characterization of a novel thermostable laccase PPLCC2 from the brown rot fungus Postia placenta MAD-698-R |
| title_short | Molecular characterization of a novel thermostable laccase PPLCC2 from the brown rot fungus Postia placenta MAD-698-R |
| title_sort | molecular characterization of a novel thermostable laccase pplcc2 from the brown rot fungus postia placenta mad 698 r |
| topic | Brown rot fungi Pichia pastoris Postia placenta Thermostable laccase |
| url | http://www.sciencedirect.com/science/article/pii/S0717345815001207 |
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