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yeast, metabolites, extraction, HPLC

Michael (2) m.hans at mx.uni-saarland.de
Fri Mar 5 17:47:37 EST 1999


hi
I am looking for information about extraction of intracellular
metabolites in the yeast S. cerevisiae. At the moment I am working with
a protocol by Gonzalez et al. 1997 (Yeast, 13:1347-1356). My problems
are:
- solution A freezes at -70°C inspite of a methanol concentration of 60%

- to concentrate the samples in the rotavapor takes at least one hour
per sample instead of 8-10 minutes (even at 120mbar and 40°C)

by the way. the analysis of extrazellular metabolites by HPLC
(AminexHPX87H) showed an unidentified substance (organic acid, sugar or
alcohol) that appears in the late exp-phase and remains constant in
stationary phase. Elution in HPLC (5mM H2SO4) takes place between
glycerol and acetate and it is detectable only by UV (210nm), but not by
RI-detector. it is not: malate, oxalacetate, lactate, citrate,
2-oxoglutarate, trehalose. what could it further be? any idea?

michael





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