The extracellular enzyme activities of Castanopsis carlesii leaf litter were evaluated during the decomposition in intermittent streams, forest floor and streams from April 2021 to July 2021 in this study. The results showed that: (1) the extracellular enzyme activities of acid phosphatase, β-Glucosidase, β-N-acetylglucosaminidase and urease were higher than those in forest floor but lower than those in streams. (2) Ecoenzymatic stoichiometry and enzyme vector features indicated that microorganisms in the three habitats were restricted by carbon and phosphorus availability in the process of leaf litter decomposition. The carbon limitation of the microbial community was higher than that in the stream and forest floor while phosphorus limitation was lower than the forest floor but higher than that in streams in the early stage of decomposition. With the progress of litter decomposition, the degree of phosphorus limitation was weakened but the carbon limitation was strengthened during the process of litter decomposition of leaf litter in subtropical intermittent streams. (3) The statistical analysis showed that air temperature greatly accounted for the variation in extracellular enzyme activities in intermittent streams, while air temperature, soil moisture content and soil temperature were more closely related to extracellular enzyme activities in forest floor. Meanwhile, the extracellular enzyme activity of stream litter was mainly affected by factors such as water temperature, stream depth and air temperature. These results provide new ideas and scientific basis for a comprehensive understanding of the material fluxes and circulation process in subtropical China.