l‐cysteine‐modified Fe3O4 nanoparticles (l‐cys@nFe3O4) were synthesized successfully and used as catalyst to activate persulfate (PS) for benzene, toluene, ethylbenzene, and xylenes (BTEX) degradation. The composite was fully characterized, and the l‐cys@nFe3O4 had more protrusions and l‐cys was combined on the surface of nFe3O4. The removals of BTEX were 78.2%, 85.1%, 85.3%, 81.2%, respectively, in PS/l‐cys@nFe3O4 system, while only 52.7% 57.8%, 60.8%, and 56.3% of BTEX removals reached under the same condition for nFe3O4 chelated with l‐cys in 48 h. Four successive cycles of BTEX degradation were completed in PS/l‐cys@nFe3O4 system. The synergistic mechanisms of BTEX degradation in PS/l‐cys@nFe3O4 system were investigated by electron paramagnetic resonance (EPR), benzoic acid (BA) probe and X‐ray photoelectron spectroscopy (XPS) tests. SFe bond in l‐cys‐Fe complexes promoted the electron transfer between nFe3O4 core and the solution, iron and iron at the interface, thereby promoting the Fe3+/Fe2+ cycle and the catalytic capacity of nFe3O4. The optimal pH of PS/l‐cys@nFe3O4 system was 3, while HCO3− and Cl− exhibited negative influences on BTEX degradation. Only 14.2%, 15.5%, 15.9%, and 15.6% BTEX had been removed in the presence of 0.12‐M PS and 8.0 g/L l‐cys@nFe3O4 under the actual groundwater condition. However, expanding the dosage of PS and l‐cys@nFe3O4 was an effective strategy to overcome the adverse effect. Practitioner Points: L‐cys@nFe3O4 were synthesized successfully and used as catalyst to activate PS for BTEX degradation.Four successive cycles of BTEX degradation were completed in PS/L‐cys@nFe3O4 system.lS―Fe bond in L‐cys@nFe3O4 promoted the electron transfer between PS and nFe3O4 core. [ABSTRACT FROM AUTHOR]