It is well established that land use change (LUC) can impact soil organic carbon (SOC) in tropical regions, but the long-term effects of LUC on soil quality and C cycling remain unclear. Here, we evaluated how LUC affects soil C cycling in the Amazon region using a 100-year observational chronosequence spanning primary forest-to-pasture conversion and subsequent secondary forest succession. We found a surprising increase in topsoil SOC concentrations 60 years following conversion, despite major losses ([ 85%) of forest-derived SOC within the first 25 years. Shifts in molecular composition of SOC, identified with diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy, occurred in tandem with a significant decline in permanganateoxidizable C (POXC) and b-glucosidase activity (per unit SOC), interpreted as a deceleration of soil C cycling after pasture grasses became the dominant source of C inputs to soil. Secondary forest succession caused rapid reversal to conditions observed under primary forest for b-glucosidase activity but not for SOC molecular composition (DRIFT spectroscopy), reflecting a long-lasting effect of LUC on soil C cycling. Our results show that rapid changes in the origin of SOC occur following deforestation with legacy effects on some indicators of C cycling (e.g. enzyme activity) but not others (e.g. molecular composition). This approach offers mechanistic