Mass spectrometry-based proteomics was applied to analyze proteins isolated from dissolved organic matter (DOM). The focal question was to identify the type and biological origin of proteins in DOM, and to describe diversity of protein origin at the level of higher taxonomic units, as well as to detect extracellular enzymes possibly important in the carbon cycle. Identified proteins were classified according to their phylogenetic origin and metabolic function using the National Center for Biotechnology Information (NCBI) protein and taxonomy database. Seventy-eight percent of the proteins in DOM from the lake but less than 50% in forest soil DOM originated from bacteria. In a deciduous forest, the number of identified proteins decreased from 75 to 28 with increasing soil depth and decreasing total soil organic carbon content. The number of identified proteins and taxonomic groups was 50% higher in winter than in summer. In spruce forest, number of proteins and taxonomic groups decreased by 50% on a plot where trees had been girdled a year before and carbohydrate transport to roots was terminated. After girdling, proteins from four taxonomic groups remained as compared to nine taxonomic groups in healthy forest. Enzymes involved in degradation of organic matter were not identified in free soil DOM. However, cellulases and laccases were found among proteins extracted from soil particles, indicating that degradation of soil organic matter takes place in biofilms on particle surfaces. These results demonstrate a novel application of proteomics to obtain a "proteomic fingerprint" of presence and activity of organisms in an ecosystem.