Identification of two novel xylanase-encoding genes (xyn5 and xyn6) from Acrophialophora nainiana and heterologous expression of xyn6 in Trichoderma reesei

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...

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Bibliographic Details
Published in:Biotechnology letters 2007-08, Vol.29 (8), p.1195-1201
Main Authors: Salles, Bruno C, Te'o, Valentino S. J, Gibbs, Moreland D, Bergquist, Peter L, Filho, Edivaldo X. F, Ximenes, Eduardo A, Nevalainen, K. M. Helena
Format: Article
Language:English
Subjects:
Acrophialophora nainiana
Amino Acid Sequence
Amino acids
Ascomycota - metabolism
Bacteria
Bacteriology
Binding Sites
Biological and medical sciences
Biotechnology
Cloning, Molecular
Densitometry - methods
Electrophoresis
Endo-1,4-beta Xylanases - chemistry
Endo-1,4-beta Xylanases - metabolism
Escherichia coli - metabolism
Fundamental and applied biological sciences. Psychology
Fungi
Gel electrophoresis
Gene Expression Regulation, Enzymologic
Gene Expression Regulation, Fungal
Genes
Genomic walking
Heterologous gene expression
Hypocrea jecorina
Introns
Microbiology
Molecular Sequence Data
Mutagenesis
Mutagenesis, Site-Directed
Nucleotide sequence
Oligonucleotides - chemistry
Open Reading Frames
Protein engineering
Proteins
Sequence analysis
Site-directed mutagenesis
Thermal stability
Trichoderma - enzymology
Trichoderma - metabolism
Trichoderma reesei
xylan
Xylanase
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Summary: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.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-007-9380-z