Standing is commonly recommended to reduce sedentary behavior in the workplace; however, constrained prolonged standing has also been linked to musculoskeletal symptoms, such as low back pain (LBP). Light physical activity breaks, such as walking, may change lumbar spine posture enough to reduce LBP during standing. This study assessed the effectiveness of inserting 5-minute walking breaks every 25 min for reducing prolonged standing-induced LBP development. Nineteen participants completed two bouts of standing lasting 2 h - one with a 5-minute walking break every 25 min and one with no breaks. Pain measures were completed throughout the trial to categorize participants as pain developers (PDs) or non-pain developers (non-PDs). Lumbar region kinematics angle and range of motion were measured continuously. In standing, 58% (11/19) of participants were PDs, compared to just 26% when walking breaks were inserted. Seventy-three percent (8/11) were categorized as non-PDs with walking breaks. Median lumbar flexion increased during walking compared to standing. Lumbar region range of motion in the coronal and transverse planes also increased during walking. The intermittent lumbar flexion may help decrease LBP during prolonged standing. These results demonstrate that walking breaks may help promote lumbar movement and reduce prolonged standing-induced LBP.