| Registro completo |
| Provedor de dados: |
BJM
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| País: |
Brazil
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| Título: |
Cloning, characterization and expression of a novel laccase gene Pclac2 from Phytophthora capsici
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| Autores: |
Feng,Bao Zhen
Li,Peiqian
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| Data: |
2014-01-01
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| Ano: |
2014
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| Palavras-chave: |
Phytophthora capsici
Laccase
Expression
Purification
Activity
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| Resumo: |
Laccases are blue copper oxidases (E.C. 1.10.3.2) that catalyze the one-electron oxidation of phenolics, aromatic amines, and other electron-rich substrates with the concomitant reduction of O2 to H2O. A novel laccase gene pclac2 and its corresponding full-length cDNA were cloned and characterized from Phytophthora capsici for the first time. The 1683 bp full-length cDNA of pclac2 encoded a mature laccase protein containing 560 amino acids preceded by a signal peptide of 23 amino acids. The deduced protein sequence of PCLAC2 showed high similarity with other known fungal laccases and contained four copper-binding conserved domains of typical laccase protein. In order to achieve a high level secretion and full activity expression of PCLAC2, expression vector pPIC9K with the Pichia pastoris expression system was used. The recombinant PCLAC2 protein was purified and showed on SDS-PAGE as a single band with an apparent molecular weight ca. 68 kDa. The high activity of purified PCLAC2, 84 U/mL, at the seventh day induced with methanol, was observed with 2,2'-azino-di-(3-ethylbenzothialozin-6-sulfonic acid) (ABTS) as substrate. The optimum pH and temperature for ABTS were 4.0 and 30 ºC, respectively . The reported data add a new piece to the knowledge about P. Capsici laccase multigene family and shed light on potential function about biotechnological and industrial applications of the individual laccase isoforms in oomycetes.
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| Tipo: |
Info:eu-repo/semantics/article
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| Idioma: |
Inglês
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| Identificador: |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000100050
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| Editor: |
Sociedade Brasileira de Microbiologia
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| Relação: |
10.1590/S1517-83822014005000021
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| Formato: |
text/html
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| Fonte: |
Brazilian Journal of Microbiology v.45 n.1 2014
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| Direitos: |
info:eu-repo/semantics/openAccess
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