RESUMEN

Two novel genes, xyn5 and xyn6, coding for family 11 xylanases, were isolated from the thermotolerant filamentous fungus, Acrophialophora nainiana, by PCR using degenerate primers. The xyn6 gene was further expressed in Trichoderma reesei. DNA sequence analysis of xyn6 revealed an open reading frame (ORF) of 708 bp, interrupted by an intron of 58 bp. The xyn6 ORF encodes a predicted protein of 236 amino acid residues. The mature recombinant XynVI protein had a molecular mass of about 19 kDa, as estimated by gel electrophoresis. Analysis of the predicted amino acid sequence of XynVI paves the way for rational protein engineering by site-directed mutagenesis aiming to increase the thermostability of the heterologously-expressed xylanase.

Asunto(s)

Ascomicetos/metabolismo , Endo-1,4-beta Xilanasas/química , Regulación Enzimológica de la Expresión Génica , Regulación Fúngica de la Expresión Génica , Trichoderma/enzimología , Secuencia de Aminoácidos , Sitios de Unión , Clonación Molecular , Densitometría/métodos , Endo-1,4-beta Xilanasas/metabolismo , Escherichia coli/metabolismo , Intrones , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Oligonucleótidos/química , Sistemas de Lectura Abierta , Trichoderma/metabolismo