Polyamide (PA) chemistry-based nanofiltration (NF) membranes have an important role in the field of seawater desalination and wastewater reclamation. Achieving an ultrathin and defect-free active layer via precisely controlled interfacial polymerization (IP) is an effective routine to improve the separation efficiencies of NF membranes. Herein, the morphologies and chemical structures of the thin-film composite (TFC) NF membranes were accurately regulated by tailoring the interfacial reaction temperature during the IP process. This strategy was achieved by controlling the temperature (−15, 5, 20, 35, and 50°) of the oil-phase solutions. The structural compositions, morphological variations, and separation features of the fabricated NF membranes were studied in detail. In addition, the formation mechanisms of the NF membranes featuring different PAs were also proposed and discussed. The temperature-assisted IP (TAIP) method greatly changed the compositions of the resultant PA membranes. A very smooth and thin PA film was obtained for the NF membranes fabricated at a low interfacial temperature; thus, a high 19.2 L m −2 h −1 bar −1 of water permeance and 97.7% of Na 2 SO 4 rejection were observed. With regard to the NF membranes obtained at a high interfacial temperature, a lower water permeance and higher salt rejection with fewer membrane defects were achieved. Impressively, the high interfacial temperature-assisted NF membranes exhibited uniform coffee-ring-like surface morphologies. The special surface-featured NF membrane showed superior separation for selected heavy metals. Rejections of 93.9%, 97.9%, and 87.7% for Cu 2+ , Mn 2+ , and Cd 2+ were observed with the optimized membrane. Three cycles of fouling tests indicated that NF membranes fabricated at low temperatures exhibited excellent antifouling behavior, whereas a high interface temperature contributed to the formation of NF membranes with high fouling tendency. This study provides an economical, facile, and universal TAIP strategy for tailoring the performances of TFC PA membranes for environmental water treatment.