Nano zero-valent iron (nZVI) particles have received much attention in environmental science and technology due to their unique electronic and chemical properties. While sulfate radical-based advanced oxidation processes (SR-AOPs) activated by nZVI show promise for mono-chlorobenzene (MCB) degradation, their efficiency is severely limited by surface oxidation of nZVI and Fe³⁺ accumulation. This study aims to enhance the nZVI/persulfate (PS) system using L-cysteine (Cys) to achieve effective MCB removal. The work involved synthesizing nZVI via borohydride reduction, followed by comprehensive characterization and batch experiments of the Cys/nZVI/PS degradation system of MCB were carried out to evaluate the key influencing factors and analyze the reaction mechanism of Cys-enhanced MCB degradation. Under optimal conditions (0.1 g/L nZVI, 3 mM PS, 0.1 mM Cys, pH 3), 92.6% of MCB was degraded within 90 min—an 18.7% improvement compared to the Cys-free system. Acidic pH promoted Fe²⁺ release and significantly enhanced degradation, while HCO₃⁻ strongly inhibited the process. Mechanistic studies revealed that sulfate radicals (SO₄^•−) played a dominant role, and Cys served as an electron shuttle that facilitated the Fe³⁺/Fe²⁺ cycle and enhanced Fe⁰ conversion, thereby sustaining PS activation. This study demonstrates that Cys effectively mitigates the limitations of nZVI/PS systems and provides valuable insights for implementing efficient SR-AOPs in treating chlorinated organic contaminants.