This study was performed to reveal the microbial community characteristics in simulated managed aquifer recharge (MAR), a natural water treatment system, under different concentrations and compositions of biodegradable dissolved organic carbon (BDOC) and further link these to the biotransformation of emerging trace organic chemicals (TOrCs). Two pairs of soil-column setups were established in the laboratory receiving synthetic feed solutions composed of different peptone/humic acid ratios and concentrations. Higher BDOC concentration resulted in lower microbial community diversity and higher relative abundance of Betaproteobacteria. Decreasing the peptone/humic acid ratio resulted in higher diversity of the community and higher relative abundances of Firmicutes, Planctomycetes, and Actinobacteria. The metabolic capabilities of microbiome involved in xenobiotics biodegradation were significantly promoted under lower BDOC concentration and higher humic acid content. Cytochrome P450 genes were also more abundant under these primary substrate conditions. Lower peptone/humic acid ratios also promoted the attenuation of most TOrCs. These results suggest that the primary substrate characterized by a more refractory character could increase the relative abundances of Firmicutes, Planctomycetes, and Actinobacteria, as well as associated cytochrome P450 genes, all of which should play important roles in the biotransformation of TOrCs in this natural treatment system.