Long, Christopher P. published the artcileHigh-resolution 13C metabolic flux analysis, Synthetic Route of 97-67-6, the main research area is Escherichia 13C metabolic flux analysis glycogen glucose ribose.
Precise quantification of metabolic pathway fluxes in biol. systems is of major importance in guiding efforts in metabolic engineering, biotechnol., microbiol., human health, and cell culture. 13C metabolic flux anal. (13C-MFA) is the predominant technique used for determining intracellular fluxes. Here, we present a protocol for 13C-MFA that incorporates recent advances in parallel labeling experiments, isotopic labeling measurements, and statistical anal., as well as best practices developed through decades of experience. The protocol is based on growing microbes in two (or more) parallel cultures with 13C-labeled glucose tracers, followed by gas chromatog.-mass spectrometry measurements of isotopic labeling of protein-bound amino acids, glycogen-bound glucose, and RNA-bound ribose. Fluxes are then estimated using software for 13C-MFA, such as Metran, followed by comprehensive statistical anal. to determine the goodness of fit and calculate confidence intervals of fluxes. The presented protocol can be completed in 4 d and quantifies metabolic fluxes with a standard deviation of =2%, a substantial improvement over previous implementations. The presented protocol is exemplified using an Escherichia coli ΔtpiA case study with full supporting data, providing a hands-on opportunity to step through a complex troubleshooting scenario. This protocol can be easily adjusted for application to eukaryotic organisms.
Nature Protocols published new progress about Computer program (Metran). 97-67-6 belongs to class alcohols-buliding-blocks, name is (S)-2-hydroxysuccinic acid, and the molecular formula is C4H6O5, Synthetic Route of 97-67-6.
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