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Metabolic network analysis of penicillium chrysogenum using 13 c‐labeled glucose
Author(s) -
Christensen Bjarke,
Nielsen Jens
Publication year - 2000
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(20000620)68:6<652::aid-bit8>3.0.co;2-j
Subject(s) - serine hydroxymethyltransferase , penicillium chrysogenum , biochemistry , glycine , serine , cytosol , chemistry , aldolase a , citrate synthase , isotopic labeling , enzyme , chromatography , biology , amino acid , organic chemistry
Using 13 C‐labeled glucose fed to a penicillin‐overproducing strain of Penicillium chrysogenum , the intracellular fluxes were quantified, and the presence of two new pathways, not previously described in this organism, is suggested. Thus, glycine was synthesized not only by serine hydroxymethyltransferase, but also by threonine aldolase. The formation of cytosolic acetyl‐CoA was found to be synthesized both via the citrate lyase‐catalyzed reaction and by degradation of the penicillin side‐chain precursor, phenoxyacetic acid. Furthermore, the experimental data indicate that the main activities of homocitrate synthase and α‐isopropylmalate synthase are located in the cytosol. All experimental data on the labeling patterns were obtained using gas chromatography–mass spectrometry, which is faster and more sensitive than the nuclear magnetic resonance methods usually applied for analysis of labeling patterns. © 2000 John Wiley & Sons, Inc. Biotechnol Bioeng 68: 652–659, 2000.