Model-independent fluxome profiling from 2H and 13C experiments for metabolic variant discrimination

We introduce a conceptually novel method for intracellular fluxome profiling from unsupervised statistical analysis of stable isotope labeling. Without a priori knowledge on the metabolic system, we identified characteristic flux fingerprints in 10 Bacillus subtilis mutants from 132 ²H and ¹³C tracer experiments. Beyond variant discrimination, independent component analysis automatically mapped several fingerprints to their metabolic determinants. The approach is flexible and paves the way to large-scale fluxome profiling of any biological system and condition.