Phenotypic effects in Saccharomyces cerevisiae after regulated expression of beta-(1,3)-glucanase from Nicotiana plumbaginifolia.
A recombinant yeast strain was constructed of which the cell wall porosity could be reversibly and conditionally modulated. This strain expresses the Nicotiana plumbaginifolia beta-(1,3)-glucanase under control of the Saccharomyces cerevisiae GAL1 promoter and the mating factor alpha 1 prepro-sequence. The following phenotypic effects were observed after expression of this enzyme: (a) expressed beta-(1,3)-glucanase is toxic to the producing yeast cells, which is reflected by a strong growth inhibition; as beta-(1,3)-glucanase could be detected only inside the cells, it seems to interfere with cell wall growth from within the cell; (b) after induction of glucanase, the recombinant strain lost up to 20% of some periplasmic enzymes, as evidenced by the release of normally periplasmic-associated invertase; (c) preliminary growth in a synthetic medium containing galactose significantly increased the transformation efficiency of the recombinant yeast strain.
Demolder
J
J
Laboratory of Molecular Biology, State University, Gent, Belgium.
De Backer
M
M
Fiers
W
W
Contreras
R
R
D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D043326:Glucan 1,3-beta-Glucosidase; D006026:Glycoside Hydrolases / Q000378:metabolism; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010641:Phenotype; D010947:Plants, Toxic; D010957:Plasmids; D011994:Recombinant Proteins / Q000378:metabolism; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014026:Nicotiana / Q000201:enzymology* / Q000235:genetics*; D043324:beta-Fructofuranosidase; D001617:beta-Glucosidase / Q000235:genetics* / Q000378:metabolism
D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D006026:Glycoside Hydrolases; D001617:beta-Glucosidase; D043324:beta-Fructofuranosidase; D043326:Glucan 1,3-beta-Glucosidase