Long-term experimental sites generally provide important information regarding how soil properties are affected by management practices. Soil microbial properties such as biomass, population, and soil enzymes have been used to predict soil quality. A wide range of enzymes has not been systematically investigated in any of the classical long-term experimental plots. This study was designed to determine effects of long-term fertilization, tillage, and crop rotations on activities of cellulase, dehydrogenase, and pytase using soil samples from historic management plots in Sanborn Field. This historic site is located on the campus of the University of Missouri at Columbia, Missouri, USA and has been under various cropping and management practices since 1888. Soil samples were collected from 0-10, 10-20, and 20-30 cm depth. It was concluded that enzymatic activities reflect cropping systems, management histories, and soil properties observed on the Sanborn Field.