Data/hora: 06/06/2024 02:26:04

Provedor de dados: 	Brazilian Journal of Microbiology	
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