A sediment control dam is an artificial structure built to prolong sedimentation in the main dam by reducing the inflow of suspended solids. These dams can affect changes in dissolved organic matter (DOM) in the water body by changing the river flow regime. The main DOM component for Yeongju Dam sediment control of the Naeseongcheon River was analyzed through 3D excitation-emission matrix (EEM) and parallel factor (PARAFAC) analyses. As a result, four humic-like components (C1~C3, C5), and three proteins, tryptophan-like components (C2, C6~C7) were detected. Among DOM components, humic-like components (autochthonous: C1, allochthonous: C2~C3) were found to be dominant during the sampling period. The total amount of DOM components and the composition ratio of each component did not show a difference for each depth according to the amount of available light (100%, 12%, and 1%). Throughout the study period, the allochthonous organic matter was continuously decomposing and converting into autochthonous organic matter; the DOM indices (fluorescence index, humification index, and freshness index) indicated the dominance of autochthonous organic matter in the river. Considering the relative abundance of cyanobacteria and that the number of bacteria cells and rotifers increased as autochthonous organic matter increased, it was suggested that the algal bloom and consequent activation of the microbial food web was affected by the composition of DOM in the water body. Research on DOM characteristics is important not only for water quality management but also for understanding the cycling of matter through microbial food web activity.