Mechanisms controlling microbial degradation of dissolved organic matter (DOM) in aquatic environments are poorly understood, although microbes are crucial to global nutrient cycling. Bacterial cell wall components may be one of the keys in understanding the presence of slowly degrading DOM in nature. We found that dominant components of bacterial cell walls (D-amino acids (D-AA), glucosamine (GluA) and diaminopimelic acid (DAPA)) comprised up to 11.4% of the dissolved organic nitrogen in 50 diverse rivers entering the Baltic Sea. Occurrence of DAPA, a characteristic component of Gram-negative (G(-)) bacteria, in the rivers suggests that G(-) bacteria rather than Gram-positive (G(+)) were the major source of the cell wall material. In laboratory studies, the degradation of whole bacterial cells, cell wall material and purified peptidoglycan was studied to characterize degradation of cell wall material by natural aquatic bacteria. Addition of whole killed G(-) and G(+) bacteria to cultures of estuarine bacteria demonstrated fragmentation and loss of cell structure of the G(+) bacteria, while the G(-) bacteria maintained an intact cell shape during the entire 69-day period. In another experiment, estuarine bacteria degraded 39-69% of GluA, D-AA and DAPA in added cell wall material of a representative G(-) bacterial species during 8 days, as compared to a 72-89% degradation of GluA, D-AA and DAPA in cell material of a G(+) bacterial species. When cultures of estuarine bacteria were enriched with purified G(-) and G(+) peptidoglycan (1 mg l(-1)), at least 49% (G(-)) and 58% (G(+)) of D-AA in the peptidoglycan was degraded. No major changes in GluA were obvious. Interpretation of the results was difficult as a portion of the purified peptidoglycan was of similar size to the bacteria and could not be differentiated from cells growing in the cultures. Addition of the purified peptidoglycan stimulated the bacterial growth, and after 6 days the cell density in the enriched cultures was 4-fold higher than in the controls. A regrowth of bacteria after addition of L-broth at 105 days caused a 50- to 75-fold increase in dissolved D-AA and GluA. Most of the D-AA and GluA were taken up during the following 10 days, indicating that cell wall constituents are dynamic compounds. Our results show that a variable portion of peptidoglycan in G(-) and G(+) bacteria can be degraded by natural bacteria, and that peptidoglycan in G(-) bacteria is more resistant to bacterial attack than that in G(+) bacteria. Thus, the presence of cell wall constituents in natural DOM may reflect the recalcitrant nature of especially G(-) peptidoglycan.