Cyclocarya paliurus is only propagated from seeds which have pronounced dormancy. Overcoming seed dormancy is an important component of efficient and cost-effective seedling production of Cyclocarya paliurus. Changes in biochemical composition and enzyme activity were investigated during dormancy release. The activities of all the studied enzymes in the stratified seeds increased significantly, compared to those in the control samples. Of the enzymes examined, the activities of protease increased the most (413.8%), followed by peroxidase (278.7%), lipase (161.0%), glucose-6-phosphate dehydrognase (149.1%) and amylase (60.6%) after 8 months of stratification. Crude fat and protein constituted the bulk of the storage reserves in mature seeds of C. paliurus. Compared with the seeds before stratification, about 45% of the starch, 46% of the protein and 11% of the crude fat were depleted during dormancy release of C. paliurus seeds, while the soluble sugar content was enhanced by 101.5% in the germinating seeds. Correlation analysis showed, during dormancy release of C. paliurus seeds, a close positive relationship between POD and G6PDH activity as well as soluble sugar content and amylase activity, while there was a significant negative relationship between storage substances and their related enzyme activities.