The targeting of Bacillus subtilis levansucrase in yeast is correlated to both the hydrophobicity of the signal peptide and the net charge of the N-terminus mature part.
We compared the ability of signal sequences from various Bacillus or yeast secreted proteins to direct Bacillus subtilis levansucrase into the secretion pathway of the yeast Saccharomyces cerevisiae. The efficiency of these sequences correlated with the overall hydrophobicity of their h-domain and was independent of their origin. Furthermore, the net charge of the proximal protein sequence downstream from the signal sequence contributed to the competence of the heterologous proteins to be secreted by yeast. Modification of this net charge allowed the protein to be translocated under the control of the yeast invertase signal sequence. Moreover, glycosylation of levansucrase did not modify significantly the fructosyl polymerase activity.
Scotti
P A
PA
Institut Jacques Monod, CNRS, Université Paris 7, France.
Praestegaard
M
M
Chambert
R
R
Petit-Glatron
M F
MF
D000135:Acid Phosphatase / Q000235:genetics; D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology; D001412:Bacillus subtilis / Q000201:enzymology*; D001692:Biological Transport; D002462:Cell Membrane / Q000201:enzymology; D003593:Cytoplasm / Q000201:enzymology; D005630:Fructans / Q000096:biosynthesis; D006026:Glycoside Hydrolases / Q000235:genetics; D006031:Glycosylation / Q000187:drug effects; D006602:Hexosyltransferases / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D021382:Protein Sorting Signals / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000378:metabolism*; D013104:Spheroplasts / Q000201:enzymology; D014415:Tunicamycin / Q000494:pharmacology; D000516:alpha-Amylases / Q000235:genetics; D043324:beta-Fructofuranosidase
D005630:Fructans; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D014415:Tunicamycin; C072599:levan; D006602:Hexosyltransferases; C021513:levansucrase; D000135:Acid Phosphatase; D006026:Glycoside Hydrolases; D000516:alpha-Amylases; D043324:beta-Fructofuranosidase