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Cloning and molecular characterization of a new fungal xylanase gene from Sclerotinia sclerotiorum S2

  • Ellouze, Olfa Elleuch (Biological Engineering Unit, National Institute of Applied Sciences and Technology (I.N.S.A.T.)) ;
  • Loukil, Sana (Biological Engineering Unit, National Institute of Applied Sciences and Technology (I.N.S.A.T.)) ;
  • Marzouki, Mohamed Nejib (Biological Engineering Unit, National Institute of Applied Sciences and Technology (I.N.S.A.T.))
  • Received : 2011.05.18
  • Accepted : 2011.07.26
  • Published : 2011.10.31
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Abstract

Sclerotinia sclerotiorum fungus has three endoxylanases induced by wheat bran. In the first part, a partial xylanase sequence gene (90 bp) was isolated by PCR corresponding to catalytic domains (${\beta}5$ and ${\beta}6$ strands of this protein). The high homology of this sequence with xylanase of Botryotinia fuckeliana has permitted in the second part to amplify the XYN1 gene. Sequence analysis of DNA and cDNA revealed an ORF of 746 bp interrupted by a 65 bp intron, thus encoding a predicted protein of 226 amino acids. The mature enzyme (20.06 kDa), is coded by 188 amino acid (pI 9.26). XYN1 belongs to G/11 glycosyl hydrolases family with a conserved catalytic domain containing $E_{86}$ and $E_{178}$ residues. Bioinformatics analysis revealed that there was no Asn-X-Ser/Thr motif required for N-linked glycosylation in the deduced sequence however, five O-glycosylation sites could intervene in the different folding of xylanses isoforms and in their secretary pathway.

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