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 (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.
Salles
Bruno C
BC
Laboratório de Enzimologia, Departamento de Biologia Celular, Universidade de Brasília, Brasilia, Brazil. cobucci@stanford.edu
Te'o
Valentino S J
VS
Gibbs
Moreland D
MD
Bergquist
Peter L
PL
Filho
Edivaldo X F
EX
Ximenes
Eduardo A
EA
Nevalainen
K M Helena
KM
D000595:Amino Acid Sequence; D001203:Ascomycota / Q000378:metabolism*; D001665:Binding Sites; D003001:Cloning, Molecular; D003720:Densitometry / Q000379:methods; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D007438:Introns; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009841:Oligonucleotides / Q000737:chemistry; D016366:Open Reading Frames; D014242:Trichoderma / Q000201:enzymology* / Q000378:metabolism
D009841:Oligonucleotides; D043364:Endo-1,4-beta Xylanases