Heterologous expression of the Bacillus pumilus endo-beta-xylanase (xynA) gene in the yeast Saccharomyces cerevisiae.
The endo-beta-xylanase-encoding gene (xynA) of Bacillus pumilus PLS was isolated from a genomic DNA library and the open reading frame (ORF) was inserted in expression vectors for the yeast Saccharomyces cerevisiae. Plasmid pFN3 harboured the xynA ORF fused to the yeast mating pheromone alpha-factor signal sequence (MFalpha1s) under the control of the alcohol dehydrogenase II gene promotor (ADH2P) and terminator (ADH2T) sequences. In plasmid pFN4, the MFalpha1S-xynA ORF was brought under the control of the phosphoglycerate kinase I gene promotor (PGK1p) and terminator (PGK1T) sequences. Autoselective, recombinant S. cerevisiae [fur1::LEU2] strains bearing pFN3 or pFN4 secreted functional endo-beta-xylanase when grown in complex medium. Enzymatic activities in the culture supernatants reached maximum levels of 8.5 nkat/ml and 4.5 nkat/ml, respectively. The temperature and pH optimum for both the bacterial and the recombinant xylanase were 58 degrees C and pH 6.2.
Nuyens
F
F
Laboratory for Food Microbiology, Katholieke Universiteit Leuven, Heverlee, Belgium. filip.nuyens@agr.kuleuven.ac.be
van Zyl
W H
WH
Iserentant
D
D
Verachtert
H
H
Michiels
C
C
D001407:Bacillus / Q000201:enzymology / Q000235:genetics*; D001709:Biotechnology / Q000379:methods; D043364:Endo-1,4-beta Xylanases; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic; D014995:Xylosidases / Q000235:genetics / Q000378:metabolism*
D014995:Xylosidases; D043364:Endo-1,4-beta Xylanases