Understanding the impact of land-use change after the conversion of natural forests is imperative for understanding soil biological activity and devising sustainable land management strategies. Nevertheless, little is known about the effect of land use alterations along the elevation on soil enzyme activity, particularly in the specific context of the Eastern Himalayan region. We selected six major land use systems (LU1 = open crop fields, LU2 = mandarin orchards, LU3 = large cardamom agroforestry, LU4 = ginger-based agroforestry, LU5 = tea plantations and LU6 = undistributed forests) across four elevational gradients (A1 = 400 - 500 m, A2 = 900 - 1000 m, A3 = 1400 - 1500 m and A4 = 1900 - 2000 m) in such a way that all six land use practices fall in every elevation gradient in the Eastern Himalayan region of West Bengal. The study revealed substantial variations in acid phosphatase activity across the land use systems, with the highest to lowest levels observed as follows: LU6 > LU5 > LU4 ≈ LU3 > LU2 > LU1. Whereas, dehydrogenase and β-glucosidase varied in the following order: LU6 > LU4 ≈ LU3> LU5 > LU2 > LU1. Altitudinal gradients had a positive influence on enzymatic response, indicating elevated enzymatic activity with increasing elevation up to A4. A redundancy analysis study showed that soil enzyme activity is driven by soil organic carbon, dissolved organic carbon, available nitrogen, and pH. The research underscores the intricate dynamics of soil enzymes, emphasizing the need to consider both land use and altitude for comprehensive soil health assessments and sustainable land management practices in the Eastern Himalayas.