title: Cloning and heterologous expression of xylanase from Pichia stipitis in Escherichia coli.
abstract: OBJECTIVE
  The main goal of this study was to characterize the xylanase (xynA) gene from Pichia stipitis NRRL Y-11543.
  
  
  RESULTS
  The xylanase gene was cloned into pUC19 in Escherichia coli DH5alphaF' and selected by growth on RBB-xylan. All functional clones contained a recombinant plasmid with an insert of 2.4 kbp, as determined by restriction mapping. The nucleotide sequence of the P. stipitis xylanase gene consisted of 1146 bp and encoded a protein of 381 amino acids with a molecular weight of 43 649 Da. The sequence contained a putative 20-amino acid N-terminal signal sequence and four N-linked glycosylation sites. The Km values for non-glycosylated and glycosylated xylanases were 1.4 mg ml-1 and 4.2 mg ml-1, respectively, and Vmax values were 0.8 and 0.082 micromol min-1 mg-1 protein, respectively.
  
  
  CONCLUSIONS
  Xylanase, a rarely found enzyme in yeast species, has been characterized in detail.
  
  
  CONCLUSIONS
  The results of this study can be used to develop better xylanase-utilizing yeast strains.
authors.affiliation: Adnan Menderes University, Gazi Bulvari, Aydin, Turkey.
authors.forename: P
authors.initials: P
authors.lastname: Basaran
authors.forename: Y D
authors.initials: YD
authors.lastname: Hang
authors.forename: N
authors.initials: N
authors.lastname: Basaran
authors.forename: R W
authors.initials: RW
authors.lastname: Worobo
mesh_terms: D000595:Amino Acid Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D010843:Pichia / Q000201:enzymology / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D043325:Xylan Endo-1,3-beta-Xylosidase; D014995:Xylosidases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D014995:Xylosidases; D043325:Xylan Endo-1,3-beta-Xylosidase