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Plant Physiol, March 2001, Vol. 125, pp. 1178-1186
Retrobiosynthetic Nuclear Magnetic Resonance Analysis of
Amino Acid Biosynthesis and Intermediary Metabolism. Metabolic Flux in
Developing Maize Kernels1
Erich
Glawischnig,2
Alfons
Gierl,
Adriana
Tomas,
Adelbert
Bacher, and
Wolfgang
Eisenreich*
Lehrstuhl für Genetik (E.G., A.G.) and Lehrstuhl für
Organische Chemie und Biochemie (A.B., W.E.), Technische
Universität München, Lichtenbergstrasse 4, 85747 Garching,
Germany; and Pioneer Hi-Bred International, 7250 NW 62nd Avenue,
Johnston, Iowa 50131-0552 (A.T.)
Information on metabolic networks could provide the basis for the
design of targets for metabolic engineering. To study metabolic flux in
cereals, developing maize (Zea mays) kernels were grown in sterile culture on medium containing
[U-13C6]glucose or
[1,2-13C2]acetate. After growth, amino acids,
lipids, and sitosterol were isolated from kernels as well as from the
cobs, and their 13C isotopomer compositions were determined
by quantitative nuclear magnetic resonance spectroscopy. The highly
specific labeling patterns were used to analyze the metabolic pathways
leading to amino acids and the triterpene on a quantitative basis. The
data show that serine is generated from phosphoglycerate, as well as from glycine. Lysine is formed entirely via the diaminopimelate pathway
and sitosterol is synthesized entirely via the mevalonate route. The
labeling data of amino acids and sitosterol were used to reconstruct
the labeling patterns of key metabolic intermediates (e.g.
acetyl-coenzyme A, pyruvate, phosphoenolpyruvate,
erythrose 4-phosphate, and Rib 5-phosphate) that revealed quantitative
information about carbon flux in the intermediary metabolism of
developing maize kernels. Exogenous acetate served as an efficient
precursor of sitosterol, as well as of amino acids of the aspartate and glutamate family; in comparison, metabolites formed in the plastidic compartments showed low acetate incorporation.
1
This research was supported by the Deutsche
Forschungsgemeinschaft (grant nos. SFB 369 and SPP 1067).
2
Present address: Department of Plant Biology, The Royal
Veterinary and Agricultural University, 40 Thorvaldsensvej, DK-1871 Frederiksberg C, Denmark.
*
Corresponding author; e-mail wolfgang.eisenreich{at}ch.tum.de;
fax 49-89-28913363.
© 2001 American Society of Plant Physiologists
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