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Biodiversity is rising rapidly on the global agenda, prompting businesses to adopt the “nature-positive” framing, expressing a commitment to combat biodiversity loss and contribute to global nature recovery goals. However, realizing these ambitions requires transformative changes in business operations, which will be challenging given the uncertainties surrounding possible strategies and pathways. A research-driven approach for business action on biodiversity is vital to prevent unintended environmental and social consequences, but there is currently no coordinated research effort on this topic. Here, we present our vision of a conceptual framework for nature positive extending beyond individual business actions, encompassing processes that influence business involvement, a spectrum of sectoral strategies, and the need for impact measurement at various scales. We utilize this framework to propose high-priority research questions where we believe collaboration between researchers, consultants, and sustainability practitioners is needed to guide effective, feasible, and equitable action to protect and restore nature.