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Critical evaluation of sampling techniques for residual glucose determination in carbon‐limited chemostat culture of Saccharomyces cerevisiae
Author(s) -
Mashego M. R.,
van Gulik W. M.,
Vinke J. L.,
Heijnen J. J.
Publication year - 2003
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.10683
Subject(s) - chemostat , fermentation , chromatography , syringe , quenching (fluorescence) , chemistry , carbon source , biochemistry , biology , fluorescence , bacteria , psychology , genetics , physics , quantum mechanics , psychiatry
Abstract In this paper, three sampling techniques for rapid quenching of cellular metabolism and subsequent separation of cells from fermentation broth are compared: (i) quick freezing of fermentation broth directly in liquid nitrogen; (ii) quenching metabolism by exposing the fermentation broth to stainless steel beads (4‐mm diameter) in a filter syringe precooled to −18°C; and (iii) withdrawal of the filtrate through a 0.45‐μm filter attached to a syringe and a needle inserted directly into the fermentor. It was concluded that use of liquid nitrogen as a quenching method to rapidly arrest cellular metabolism, for quantitative analysis of extracellular glucose, is not a very reliable method and that the filter syringe steel beads work very well. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 395–399, 2003.