We studied the effects of a bacterium (Pseudomonas chlororaphis) and a bactivorous protozoan (Uronema sp.) on transformations of labile dissolved organic carbon (DOC). In 36-day time series experiments, bacteria were grown on glucose both with and without protozoa. We measured bulk organic carbon pools and used electrospray ionization mass spectrometry to characterize dissolved organic matter on a molecular level. Bacteria rapidly utilized glucose, depleting it to nondetectable levels and producing new DOC compounds of higher molecular weight within 2 days. Some of these new compounds, representing 3 to 5% of the initial glucose-C, were refractory and persisted for over a month. Other new compounds were produced and subsequently used by bacteria during the lag and exponential growth phases, pointing to a dynamic cycling of organic compounds. Grazers caused a temporary spike in the DOC concentration consisting of labile compounds subsequently utilized by the bacteria. Grazing did not increase the complexity of the DOC pool already established by the bacteria but did continually decrease the particulate organic carbon pool and expedited the conversion of glucose-C to CO2. After 36 days, 29% of initial glucose-C remained in pure bacteria cultures, while only 6% remained in cultures where a grazer was present. In this study the bacteria were the primary shapers of the complex DOC continuum, suggesting higher trophic levels possibly have less of an impact on the qualitative composition of DOC than previously assumed.