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Metabolites 2016, 6(3), 21; doi:10.3390/metabo6030021

Decoding Biosynthetic Pathways in Plants by Pulse-Chase Strategies Using 13CO2 as a Universal Tracer

Lehrstuhl für Biochemie, Technische Universität München, 85748 Garching, Germany
Dedicated to Dr. Karl-Heinz Michl on the occasion of his 90th birthday
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Author to whom correspondence should be addressed.
Academic Editor: Peter Meikle
Received: 24 May 2016 / Revised: 3 July 2016 / Accepted: 4 July 2016 / Published: 14 July 2016
(This article belongs to the Special Issue Carbon Metabolism)

Abstract

13CO2 pulse-chase experiments monitored by high-resolution NMR spectroscopy and mass spectrometry can provide 13C-isotopologue compositions in biosynthetic products. Experiments with a variety of plant species have documented that the isotopologue profiles generated with 13CO2 pulse-chase labeling are directly comparable to those that can be generated by the application of [U-13C6]glucose to aseptically growing plants. However, the application of the 13CO2 labeling technology is not subject to the experimental limitations that one has to take into account for experiments with [U-13C6]glucose and can be applied to plants growing under physiological conditions, even in the field. In practical terms, the results of biosynthetic studies with 13CO2 consist of the detection of pairs, triples and occasionally quadruples of 13C atoms that have been jointly contributed to the target metabolite, at an abundance that is well above the stochastic occurrence of such multiples. Notably, the connectivities of jointly transferred 13C multiples can have undergone modification by skeletal rearrangements that can be diagnosed from the isotopologue data. As shown by the examples presented in this review article, the approach turns out to be powerful in decoding the carbon topology of even complex biosynthetic pathways. View Full-Text
Keywords: biosynthesis; metabolism; isotopologue profiling; 13CO2; flux analysis biosynthesis; metabolism; isotopologue profiling; 13CO2; flux analysis
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Bacher, A.; Chen, F.; Eisenreich, W. Decoding Biosynthetic Pathways in Plants by Pulse-Chase Strategies Using 13CO2 as a Universal Tracer. Metabolites 2016, 6, 21.

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