Data/hora: 15/06/2024 20:17:23

Provedor de dados: | 58
País: | Brazil
Título: | Cloning, characterization and expression of a novel laccase gene Pclac2 from Phytophthora capsici
Autores: | Feng,Bao Zhen; Li,Peiqian.
Data: | 2014-01-01
Ano: | 2014
Palavras-chave: | Phytophthora capsici; Laccase; Expression; Purification; Activity.
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.
Tipo: | Info:eu-repo/semantics/article
Idioma: | Inglês
Identificador: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000100050
Editor: | Sociedade Brasileira de Microbiologia
Relação: | 10.1590/S1517-83822014005000021
Formato: | text/html
Fonte: | Brazilian Journal of Microbiology  v.45 n.1 2014
Fonte: | info:eu-repo/semantics/openAccess