Microbial communities will play a critical role in determining arctic ecosystem response to warming due to global climate change since decomposition, which is responsible for both CO₂ evolution and plant nutrient availability, is a microbially-mediated process. Microorganisms living in tundra soil have access to two broad categories of carbon compounds via decomposition: complex polymers such as cellulose, lignin, protein and soil organic matter, and simple monomers such as glucose, phenolics and amino acids. Thermodynamically, labile substrates are easier to degrade because the activation energy of the reaction is lower than for more recalcitrant polymers. Thus, metabolic processes involving simple compounds are more likely under frozen conditions during the cold season. To test this theory, we incubated soils collected at Toolik Lake, Alaska before and after snowmelt and freeze-up with a variety of ¹³C-labelled substrates to track microbial carbon use during cold- and growing-season conditions. Surprisingly, tussock soils respired more protein-derived carbon under frozen pre-thaw conditions than late growing-season conditions. Organic shrub soils respired more glutamic acid-derived carbon under all conditions. Since nitrogen is more abundant in shrub than tussock soils, microbes in the shrub soil can afford to respire glutamic acid as an energy source rather than incorporate it into microbial biomass as a nitrogen source. This study indicates that winter substrate use may not follow predictions made strictly on a thermodynamic basis and hints at the complex nature of cold-tolerant microorganisms.