Novel thin-film nanocomposite nanofiltration ((TFN)-NF) membrane was successfully fabricated by adding MoS2 functionalized with cysteine in a polysulfone (PSf) layer and the active layer to enhance separation performance in water and alcohol. A novel TFN-NF membrane was prepared through interfacial polymerization (IP), which reacts with an aqueous DETA solution containing MoS2-cys and a TMC organic solution onto the PSF/MoS2-cys support layer. A PSf support layer was incorporated with MoS2-cys, which enhanced the membrane permeability due to increasing porosity and hydrophilicity. Furthermore, MoS2 is essential in forming thin films and Turing structures of the NF membrane, resulting in enhanced permeability for water and alcohol. The prepared TFN-NF membrane has shown a notable permeability of 7.9 LMH/bar and a rejection rate of 99% for MgSO4. It also demonstrated excellent fouling resistance after three chemical cleaning cycles and long-term running. The TFN-NF membrane with MoS2 showed improved permeability for water and alcohol solvents, such as methanol and ethanol, of 5.5, 1.2, and 0.48 LMH/bar, respectively. This work would present a valuable strategy for preparing a TFN-NF membrane to retrieve alcohol solvents.