This study investigated the acute impact of heavy metals on activated sludge with respect to the amount properties of biopolymers and other solvable microbiological products (SMPs) released from the sludge. Ten heavy metals were selected for the evaluation. Under the experimental conditions, exposing activated sludge to different metals led to an increase in SMPs, with a more significant increase in nitrogenous organics than in carbonaceous ones, where Hg2+, Ag+, Cu2+, and Cr6+ led to the highest increase in SMP species, while Cd2+, Ni2+, Mn2+, Pb2+, and Co2+ caused limited increase in the middle and small SMP molecules, and Zn2+ and Cr3+ resulted in a decrease in SMP content. To probe the molecular impact of heavy metals and the association between cellular stress and SMP formation, the toxicity of heavy metals was evaluated using a toxicogenomics assay. Based on a correlation analysis between the increase in SMP and the molecular toxicity index-transcriptional effect level index (TELI) of different genes under corresponding stress conditions, eight genes demonstrated a strong correlation with SMP properties and were pre-assumed to have the most significant influence on the increment in SMPs. We further validated the correlation equation established to predict SMP production based on the molecular disturbance of the eight key biomarkers, using arsenic As3+ and vanadium V5+ as tests, and by quantifying the amount of SMPs released from the activated sludge under the influence of these metals using a TELI-derived equation. In addition, the heavy metals that generated greater amounts of reactive oxygen species also caused larger increases in SMPs.