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Aim: Low‐molecular‐weight organic substances (LMWOSs) are at the nexus between micro‐organisms, plant roots, detritus, and the soil mineral matrix. The nominal oxidation state of carbon (NOSC) has been suggested as a potential parameter for modelling microbial uptake rates of LMWOSs and the efficiency of carbon incorporation into new biomass. Methods and Results: In this study, we assessed the role of compound class and oxidation state on uptake kinetics and substrate‐specific carbon use efficiency (SUE) during the growth of three model soil micro‐organisms, a fungal isolate (Penicillium spinulosum) and two bacterial isolates (Paraburkholderia solitsugae, and Ralstonia pickettii). Isolates were chosen that spanned a growth rate gradient (0.046–0.316 h−1) in media containing 34 common LMWOSs at realistically low initial concentrations (25 μM each). Clustered, co‐utilization of LMWOSs occurred for all three organisms. Potential trends (p < 0.05) for early utilization of more oxidized substrates were present for the two bacterial isolates (P. solitsugae and R. pickettii), but high variability (R2 < 0.15) and a small effect of NOSC indicate these relationships are not useful for prediction. The SUEs of selected substrates ranged from 0.16 to 0.99 and there was no observed relationship between NOSC and SUE. Conclusion: Our results do not provide compelling population‐level support for NOSC as a predictive tool for either uptake kinetics or the efficiency of use of LMWOS in soil solution. Significance and Impact of the Study: Metabolic strategies of organisms are likely more important than chemical identity in determining LMWOS cycling in soils. Previous community‐level observations may be biased towards fast‐responding bacterial community members. [ABSTRACT FROM AUTHOR]