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Metabolic network analysis of lysine producing Corynebacterium glutamicum at a miniaturized scale
Author(s) -
Wittmann Christoph,
Kim Hyung Min,
Heinzle Elmar
Publication year - 2004
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.20103
Subject(s) - corynebacterium glutamicum , metabolic flux analysis , flux (metallurgy) , pentose phosphate pathway , biochemistry , lysine , metabolic engineering , chemistry , microtiter plate , metabolic pathway , metabolic network , strain (injury) , chromatography , biology , metabolism , amino acid , glycolysis , enzyme , organic chemistry , gene , anatomy
Abstract We present a straightforward approach comprising 13 C tracer experiments at 200‐μL volume in 96‐well microtiter plates with on‐line measurement of dissolved oxygen for quantitative high‐throughput metabolic network analysis at a miniaturized scale. This method was successfully applied for cultivation and 13 C metabolic flux analysis of two mutants of lysine producing Corynebacterium glutamicum (ATCC 13287 and ATCC 21543). Microtiter‐plate cultivations showed excellent accordance in kinetics and stoichiometry of growth and product formation as well as in intracellular flux distributions as compared with parallel shake‐flask experiments. These cultivations further allowed clear identification of strain‐specific flux differences such as increased flux toward lysine, increased flux through the pentose phosphate pathway (PPP), decreased flux through the tricarboxylic (TCA) cycle, and increased dihydroxyacetone formation in C. glutamicum ATCC 21543 compared with ATCC 13287. The present approach has strong potential for broad quantitative screening of metabolic network activities, especially those involving high‐cost tracer substrates. © 2004 Wiley Periodicals, Inc.