Extreme drought can strongly impact belowground communities and biogeochemical processes, including soil microbial community composition and extracellular enzyme activities (EEAs), which are considered key agents in ecosystem carbon (C) and nutrient cycling. However, our understanding of how seasonal timing of drought during the growing season affects soil microbial communities and their activity remains notably poor. In this study, we investigated the responses of soil physicochemical properties, EEAs, and bacterial community composition to extreme-duration drought imposed in the early-, mid-, or late-stages of the growing season in a semiarid grassland ecosystem in Inner Mongolia, China. Compared with the ambient control, the activities of C-, nitrogen (N)-, and phosphorus (P)-acquisition enzymes were significantly decreased in the mid- and/or late-stages of drought. Bacterial community diversity also significantly decreased in the mid- and late-stage drought treatments. Soil water content was the most important factor explaining changes in soil EEAs and bacterial community composition. At the end of the growing season, the activities of C-, N-, and P-acquisition enzymes had mostly recovered, while the bacterial community diversity in the mid- and late-stage drought treatments was still lower than the ambient control. Overall, our study demonstrates that the effects of extreme drought on soil EEAs and bacterial community composition depend on the timing of drought. Our results highlight that understanding the effects of extreme-duration drought at different stages of the growing season may play a vital role in predicting the responses of belowground function to global changes in grassland ecosystems.