Over the last 20 years, the United States Agency for International Development (USAID) has supported conservation enterprises in an estimated quarter of its biodiversity programs to create incentives for stakeholders to reduce threats to biodiversity. These enterprises included timber and non-timber forest products, and ecotourism services, among others. While reviews and guidance materials are available on the conditions needed to establish conservation enterprises, there is less information on what it takes to sustain enterprises and achieve longer-term conservation outcomes. USAID’s Conservation Enterprises Learning Group developed a generic theory of change that outlines commonly held assumptions about the path from supporting enterprises to achieving biodiversity conservation. The theory of change provided a framework for cross-site learning for past and current enterprise strategies supported by USAID. The learning group generated insights to help practitioners improve the design and implementation of this strategy, including using a monitoring and learning framework for practitioners to measure outcomes from their actions over time and share lessons with the conservation community. It is our hope that this framework will be useful for conservation organizations, funders, and researchers who are considering using, supporting, and assessing enterprises so that they can more effectively support conservation.