Relating growth dynamics and glucoamylase excretion of individual Saccharomyces cerevisiae cells.
We have developed a novel flow cytometric procedure that allows determinations of properties of protein excretion in the growth medium on a cell-by-cell basis in Saccharomyces cerevisiae. The procedure is based on labelling of a periplasmically secreted protein with antibodies conjugated to a fluorescent marker such as fluorescein isothiocyanate (FITC). The staining conditions did not perturb cell growth after resuspension of stained cells in growth medium. Decrease in fluorescence was found to correlate with excretion of glucoamylase into the growth medium. The analysis of the staining pattern over time provides information on the behaviour of individual cells belonging to different cell-cycle phases and can be used to calculate the specific excretion rate of the overall population.
Porro
D
D
Department of Biotechnology and Biosciences, Università degli Studi di Milano-Bicocca, P.zza della Scienza N degrees 2, 20126, Milan, Italy. danilo.porro@unimib.it
Venturini
M
M
Brambilla
L
L
Alberghina
L
L
Vanoni
M
M
D003470:Culture Media; D005434:Flow Cytometry / Q000379:methods*; D005087:Glucan 1,4-alpha-Glucosidase / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000254:growth & development*
D003470:Culture Media; D005087:Glucan 1,4-alpha-Glucosidase