Eight constructed wetland microcosm (CWM) units have been designed using three macrophytes for domestic wastewater treatment. The main aim of this study is to evaluate enzyme activities with respect to time and soil depth and their correlation with removal efficiency of pollutants within different CWM units. The findings of this study show that the activity of enzymes and pollutants removal efficiency vary to a great extent on the soil depth, time of the sampling and type of pollutants. The correlation between removal of soluble reactive phosphorus and total phosphorus was significant with phosphatase activity in most of the CWM units. Activity of urease and NH4+-N removal was positively correlated with significant positive correlation in CWM units planted with Phragmites karka, and Pistia stratiotes (Ph + Pi) and Typha latifolia, Phragmites karka and Pistia stratiotes (T + Ph + Pi). Urease activity was found to be both positively and negatively correlated with respect to removal of NO3--N and NO2--N in different CWM units. Dehydrogenase activity showed negative correlation with respect to biological oxygen demand (BOD) removal except in CWM units with Ph + Pi and T + Ph + Pi. Similarly, a moderate positive and negative correlation exists between fluorescein diacetate hydrolysis and BOD removal. Removal of BOD and microbial biomass carbon (MBC) was negatively correlated with each other in most of the CWM units. With respect to vertical variation, the top layer of CWM units expressed significantly higher activity of extracellular enzymes and were significantly different from the deeper layer. CWM units exhibited significant variations in enzyme activity with respect to time. Copyright © 2020 Elsevier Ltd. All rights reserved.