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In vivo analysis of metabolic dynamics in Saccharomyces cerevisiae : I. Experimental observations
Author(s) -
Theobald Uwe,
Mailinger Werner,
Baltes Michael,
Rizzi Manfred,
Reuss Matthias
Publication year - 1997
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/(sici)1097-0290(19970720)55:2<305::aid-bit8>3.0.co;2-m
Subject(s) - glycolysis , biochemistry , saccharomyces cerevisiae , nad+ kinase , metabolite , extracellular , in vivo , yeast , chemistry , metabolism , enzyme , biology , microbiology and biotechnology
The goal of this work was to obtain rapid sampling technique to measure transient metabolites in vivo. First, a pulse of glucose was added to a culture of the yeast Saccharomyces cerevisiae growing aerobically under glucose limitation. Next, samples were removed at 2 to 5 s intervals and quenched using methods that depend on the metabolite measured. Extracellular glucose, excreted products, as well as glycolytic intermediates (G6P, F6P, FBP, GAP, 3‐PG, PEP, Pyr) and cometabolites (ATP, ADP, AMP, NAD + , NADH) were measured using enzymatic or HPLC methods. Significant differences between the adenine nucleotide concentrations in the cytoplasm and mitochondria indicated the importance of compartmentation for the regulation of the glycolysis. Changes in the intra‐ and extracellular levels of metabolites confirmed that glycolysis is regulated on a time scale of seconds. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55 : 305–316, 1997.