Leather manufacturing is increasingly prioritizing environmentally friendly processes, emphasizing clean production to reduce environmental impacts [1-3]. The present work was focused to explore the application of an α-amylase/neutral protease system (ANS) in a simplified one-step process for unhairing, fiber opening, and bating to replace the traditional, chemically beamhouse of lime-sulfide system (LSS). With response surface methodology (RSM), a mathematical model was established to optimize operational conditions, with the concentrations of 0.3 wt.% α-amylase and 0.5 wt.% neutral protease at 28.4℃ for 16.6 hours. The effectiveness of the process on unhairing and fiber opening was studied through scanning electron microscopy (SEM), and the impact on bating was evaluated by the removal rates of carbohydrate and proteoglycan. The leather produced using the optimized ANS exhibited comparable physical properties to those traditionally processed, with higher hydrothermal shrinkage temperature and better softness. Environmentally, the optimized ANS process achieved significant reductions in pollutants, more than 90% of chemical oxygen demand (COD), NH₃-N, and Cl^-, and 73.91% of total solids (TS) by. An economic analysis further revealed a direct cost savings of 30.98% with the ANS of that with the LSS, alongside indirect benefits of enhanced production efficiency and simplified wastewater treatment. Notably, the one-step enzymatic beamhouse substantially decreases the electricity and water usage, potentially reducing the greenhouse gas emissions by 44.6%. The ANS is proposed to be a sustainable and cost-effective alternative for leather manufacturing with environmentally friendly practices.