Abstract
C-Isotopologue profiling is a powerful tool to determine on a quantitative basis the biosynthetic origin of carotenoids in microorganisms. To this aim, the carotenoid-producing microorganism is grown in medium containing 13C-labeled glucose. After growth, the 13C-isotopologue distribution in a given biosynthetic carotenoid is determined by quantitative NMR spectroscopy. The labeling pattern provides a fingerprint of processes involved in the metabolism of glucose and the formation of the carotenoid. For example, the 13C-profile shows whether the isoprenoid precursors, isopentenyl diphosphate and dimethylallyl diphosphate, are formed by the mevalonate or the non-mevalonate pathway. The labeling data also specify the pathways of glucose utilization, e.g., via the Entner-Doudoroff pathway or glycolysis. The method is exemplified with the analysis of zeaxanthin biosynthesis in the Alphaproteobacterium, Paracoccus zeaxanthinifaciens.
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References
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Eylert, E., Bacher, A., Eisenreich, W. (2012). NMR-Based Isotopologue Profiling of Microbial Carotenoids. In: Barredo, JL. (eds) Microbial Carotenoids from Bacteria and Microalgae. Methods in Molecular Biology, vol 892. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-879-5_20
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DOI: https://doi.org/10.1007/978-1-61779-879-5_20
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-61779-879-5
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