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Reciprocal 13 C‐Labeling: A Method for Investigating the Catabolism of Cosubstrates
Author(s) -
Christensen Bjarke,
Nielsen Jens
Publication year - 2002
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp010152d
Subject(s) - penicillium chrysogenum , amino acid , catabolism , chemistry , cofactor , biosynthesis , biochemistry , substrate (aquarium) , carbon fibers , metabolism , enzyme , biology , ecology , materials science , composite number , composite material
The principle of reciprocal labeling is to use a uniformly 13 C‐labeled substrate as the primary carbon source and a naturally labeled cosubstrate. Metabolites derived from a naturally labeled cosubstrate, in this case amino acids, can then be identified by their relatively lower content of 13 C, and information on the degradation pathway can be deduced. The technique is based on GC–MS measurements of amino acid labeling patterns, making the technique well suited for investigating the relative importance of amino acid biosynthesis and amino acid uptake from the medium, as the 13 C content of the amino acids incorporated into biomass is a direct measure of the amino acid biosyntheses. The technique is illustrated by the investigation of the degradation of phenoxyacetic acid, a medium component that is essential for production of penicillin V by Penicillium chrysogenum. Glucose was used as the uniformly labeled primary carbon source.