Study on agglutinating factors from flocculent Saccharomyces cerevisiae strains.
The lectin-like theory suggest that yeast flocculation is mediated by an aggregating lectinic factor. In this study we isolated an agglutinating factor, which corresponds to lectin, from whole cells by treating the flocculent wild-type Saccharomyces cerevisiae NCYC 625 strain and its weakly flocculent mutant [rho degrees ] with EDTA and two non-ionic surfactants (Hecameg and HTAC). The dialysed crude extracts obtained in this way agglutinated erythrocytes and this hemagglutination was specifically inhibited by mannose and mannose derivatives. However, SDS-PAGE profiles showed that the three reagents had different effects on the yeast cells. The non-ionic surfactants appeared to be the most efficient, as their extracts possessed the highest specific agglutinating activity. The products released by the wild-type strain presented a higher specific agglutinating activity than those released by the [rho degrees ] mutant. Purification of the agglutinating factor from extracts of both strains by affinity chromatography revealed two active bands of relative mass of 26 and 47 kDa on SDS-PAGE. Mass spectrometry analysis by MALDI-TOF, identified a 26 kDa band as the triose phosphate isomerase (TPI) whereas a 47 kDa band was identical to enolase. Edman degradation showed that the N-terminal sequences of these proteins were similar to TPI and enolase, respectively. The difference in the flocculation behaviour of the two strains is due to changes in the protein composition of the cell wall and in the protein structure involved in cell-cell recognition.
Ngondi-Ekome
J
J
Laboratoire de Chimie Physique et Microbiologie pour l'Environnement, UMR-CNRS 7564, UHP Nancy 1, Faculté de Pharmacie, Biochimie Microbienne, 5, rue Albert-Lebrun, B.P. 403-54001 Nancy cedex, France.
Thiebault
F
F
Strub
J-M
JM
Van Dorsselaer
A
A
Bonaly
R
R
Contino-Pepin
C
C
Wathier
M
M
Pucci
B
B
Coulon
J
J
D000373:Agglutinins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D002219:Carbamates; D002473:Cell Wall / Q000737:chemistry / Q000378:metabolism; D003902:Detergents; D004492:Edetic Acid; D004591:Electrophoresis, Polyacrylamide Gel; D005427:Flocculation; D005960:Glucosides; D006386:Hemagglutination Tests; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008766:Methylmannosides; D008969:Molecular Sequence Data; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000373:Agglutinins; D002219:Carbamates; D003902:Detergents; D005960:Glucosides; D037102:Lectins; D008766:Methylmannosides; C060539:6-O-(N-heptylcarbamoyl)methylglucoside; D004492:Edetic Acid