Purpose <p>The purpose of this study is to compare the functional outcomes and donor site morbidities after anterior cruciate ligament reconstruction (ACLR) using peroneus longus (PL) tendon versus hamstring (HST) tendon autografts.</p> Methods <p>The PL tendon autograft was used for ACLR in 36 patients, and in another group, ACLR was performed using the HST tendon autograft in 35 patients between September 2022 and April 2023. The knee functional outcomes were evaluated using the International Knee Documentation Committee (IKDC) and the Lysholm scores at preoperative and at 18 months following ACLR. In addition, the autograft diameter was measured intraoperatively in both groups. Ankle joint donor site morbidities were estimated using the American Orthopedic Foot and Ankle Score (AOFAS) in the PL autograft group.</p> Results <p>A total of 71 patients, who underwent ACLR, were assessed with a minimum follow-up of 18 months (range 18–20 months). The diameter of the PL tendon autograft was significantly greater than that of the HST tendon autograft (<i>P</i> &lt; 0.001). No significant differences were found in the functional outcomes between both groups at 18 months follow-up. Evaluation of the AOFAS showed no significant ankle joint dysfunction in the PL tendon autograft group.</p> Conclusions <p>PL tendon autograft can be used as a safe and effective autograft choice for ACLR with excellent functional outcomes comparable to HST tendon autograft and minimal donor site morbidity.</p> Level of evidence <p>Level II, Prospective randomized comparative study.</p>